Mlk inhibitors and methods of use

ABSTRACT

Provided are compounds having an inhibitory effect on Mixed Lineage Kinases. Also provided are pharmaceutical compositions, methods of preparing the compounds, synthetic intermediates, and methods of using the compounds, independently or in combination with other therapeutic agents, for treating diseases and conditions which are affected by Mixed Lineage Kinase inhibition. Also provided are methods of treatment of neuropsychiatric disorders which comprise the inhibition of Mixed Lineage Kinases.

This application claims the benefit of U.S. Provisional Applications No.61/117,950, filed Nov. 25, 2008, No. 61/148,755 filed Jan. 30, 2009, andNo. 61/148,778 filed Jan. 30, 2009, all of which are incorporated byreference as if written herein in their entireties.

This research has been funded in part by NIH Grant No: NIH P01 3MH64570.

Mammalian protein kinases are involved in the regulation of importantcellular functions. Due to the fact that dysfunctions in protein kinaseactivity have been associated with several diseases and disorders,protein kinases are targets for drug development.

Mixed lineage kinases (MLKs) are MAPK kinase kinases that target JNK andp38 MAPK for activation in response to diverse stimuli that stresscells. As a result, the MLKs regulate a broad range of cellularprocesses. MLK3 is the most widely expressed MLK family member and ispresent in neurons. It is activated by GTPases of the Ras superfamily,such as Cdc42 and Rac, which trigger protein dimerization via a leucinezipper interface, resulting in auto-phosphorylation at Thr277 and Ser281within the protein activation loop and subsequent activation of theenzyme.

Preclinical studies of the mixed lineage kinase (MLK) inhibitor CEP1347have shown that this agent can protect neurons against a considerablerange of insults, including exposure to the Alzheimer's peptide, Aβ.Studies using the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine model ofParkinsonism have demonstrated the efficacy of CEP1347 in treating motordeficits and neuronal degeneration, and CEP1347-mediated neuroprotectionhas also been observed in an in vitro model for Parkinson's Disease,using methamphetamine-exposed human mesencephalic-derived neurons. Thisfinding suggests that CEP1347 might also be protective in the context ofneurologic complications such as HIV-associated dementia (HAD). In fact,Bodner et al. have shown that CEP1347 can protect primary rathippocampal neurons as well as dorsal root ganglion neurons from theotherwise lethal effects of exposure to HIV-1 gp120. It has beendetermined that CEP1347 mediates this effect by inhibiting the activityof the mixed lineage kinase (MLK) family.

Maggirwar et al. recently examined the effect of Tat and gp120 on MLK3.Tat and gp120 were shown to induce autophosphorylation of MLK3 inprimary rat neurons and this was abolished by the addition of CEP1347.These studies suggest that the normal function of MLK3 is compromised byHIV-1 neurotoxins, resulting in the downstream signaling events thatresult in neuronal death and monocyte activation (with release ofinflammatory cytokines). Most recently, Eggert et al. have demonstratedthat CEP1347 is neuroprotective in an in vivo model of HIV-1 infection,reversing microglial activation and restoring normal synapticarchitecture, as well as restoring macrophage secretory profiles to atrophic vs. toxic phenotype in response to HIV-1 infection. Eggert, D.,Gorantla, S., Poluekova, L., Dou, H., Schifitto, G., Maggirwar, S. B.,Dewhurst, S., Gelbard, H. A. and H. E. Gendelman: “NeuroprotectiveActivities of CEP-1347 in Models of HIV-1 Encephalitis,” J. Immunol. (inpress).]

Recently, MLK3 has been shown to drive the production of the HIV virus.As a result, several lines of evidence now support that an inhibitor ofMLK3 could serve as a treatment for numerous neurological conditions,including neuroAIDS. CEP1347 does not have ideal pharmacokineticproperties, which could potentially affect its ability to gain entry tothe CNS. Other small molecule MLK3 inhibitors are needed that haveimproved pharmacokinetic and brain penetrating properties.

An inhibitor of MLK3 could also find use in the treatment ofpsychological disorders. Depression is a complex disease that has amultifactorial etiology. This may include genetic factors, changes innormal neuronal signaling, and reduced levels of certain neurotrophins(such as brain-derived neurotrophic factor, BDNF) within particularregions of the brain (Krishnan, V., and E. J. Nestler. 2008. Nature455:894-902). Treatments for depression include drugs such as SSRIs, aswell as cognitive and behavioral therapy (“talk therapy”) and otherinventions such as exercise. Interestingly, SSRIs and exercise share thecommon property that they promote neurogenesis; this is thought to berelated to their anti-depressive effects because of effects on neuronalplasticity and remodeling (Krishnan, supra).

Pharmacologic blockade of mixed lineage kinase 3 (MLK3) has been shownto result in activation of neurotrophin-mediated signaling pathways, andincreased expression of neurotrophin receptors—resulting in enhancedresponsiveness to endogenous neurotrophins, including BDNF (Wang, L. H.,A. J. Paden, and E. M. Johnson, Jr. 2005. J Pharmacol Exp Ther312:1007-19). MLK3 inhibitors have also been shown to increaseproduction of BDNF itself (Conforti, P. et al. 2008. Mol Cell Neurosci39:1-7).

Combined treatment with SSRIs and MLK3 inhibitors could result in thesynergistic promotion of neurogenesis, due to theneurotrophin-sensitizing effects of MLK3 inhibitors and their ability todirectly upregulate BDNF (Wang and Conforti, supra). Increase of thetherapeutic effectiveness of SSRIs (and possibly talk therapy andexercise also) could also result if the compounds were coadministered.

Exposure to MLK3 inhibitors may also compensate for lowered BDNF levelsin hippocampus of persons with depression, thereby alleviatingdepression (based on the “BDNF hypothesis”) (Krishnan, supra).

Disclosed herein are compounds having an inhibitory effect on MLK1,MLK2, and MLK3. In a related aspect, also disclosed herein are compoundsof Formula I as described below. Thus, provided herein are novelcompounds that can be used for therapeutic methods involving modulationof MLKs. Also provided are pharmaceutical compositions, methods ofpreparing the compounds, synthetic intermediates, and methods of usingthe compounds, independently or in combination with other therapeuticagents, for treating diseases and conditions affected by MLK inhibition.

In one aspect, the present invention provides for compounds of FormulaI:

wherein:

dashed lines indicate that a second bond may alternatively be present orabsent;

X₁ is chosen from CH and N;

X₂ is chosen from C and N;

Y₁ is —(CR_(6a)R_(6b))_(m)—Z₁—(CR_(7a)R_(7b))_(n)—;

Y₂ is —(CR_(8a)R_(8b))_(p)—Z₂—(CR_(9a)R_(9b))_(q)—;

Y₃ is —(CR_(10a)R_(10b))_(r)—Z₃—(CR_(11a)R_(11b))_(s)—;

Y₄ is —(CH₂)_(t)—Z₄—;

Z₁, Z₂, and Z₃, are each independently chosen from a bond, O, S, S(O),S(O)₂, N(R₁₂), C(O), C(O)N(R₁₂), N(R₁₂)C(O), S(O)₂N(R₁₂), andN(R₁₂)S(O)₂;

Z₄ is chosen from a bond, O, and N; m, n, p, q, r, and s are eachindependently an integer from 0 to 6;

t is an integer from 0 to 2;

R₁, R₂, and R₃ are independently chosen from hydrogen, halo, loweralkyl, lower alkenyl, lower alkynyl, lower haloalkyl, lower cycloalkyl,heterocycloalkyl, aryl, heteroaryl, acyl, amido, amino, alkoxy, hydroxy,cyano, and nitro, any of which may be optionally substituted; or R₁ andR₂ may each additionally be heteroalkyl, and may be joined together suchthat R₁ and R₂ together form an alkylene, alkenylene, or heteroalkylbridge comprising from 3 to 5 atoms, which may be optionallysubstituted;

R₄ is chosen from hydrogen, (O), (S), halogen, hydroxy, cyano, nitro,lower alkyl, lower alkenyl, lower alkynyl, lower cycloalkyl, lowercycloalkyloxy, lower thioalkoxy, lower heterocycloalkyl, aryl, loweraralkyl, lower heteroaryl, lower heteroaralkyl, amido, acyl, amino, andlower alkoxy, any of which may be optionally substituted; or R₃ and R₄may each additionally be heteroalkyl, and may be joined together suchthat R₁ and R₂ together form an alkylene, alkenylene, or heteroalkylbridge comprising from 3 to 5 atoms, which may be optionallysubstituted;

R₅ and R₁₃ are each independently chosen from hydrogen, halogen,hydroxy, cyano, nitro, lower alkyl, lower alkene, lower alkyne, loweraryl, lower arylalkyl, lower cycloalkyl, lower cycloalkylalkyl, lowerheteroaryl, lower heteroarylalkyl, lower heterocycloalkyl, lowerheterocycloalkylalkyl, and lower alkoxy, any of which may be optionallysubstituted; and additionally, R₁₃ and R₃ may be joined together to forma lower spiro-cycloalkyl or spiro-phenyl comprising from 3 to 6 atoms,which may be optionally substituted;

R_(6a), R_(6b), R_(7a), R_(7b), R_(8a), R_(8b), R_(9a), R_(9b), R_(10a),R_(10b), R_(11a), R_(11b), and R₁₂ are each independently chosen from abond, hydrogen, halogen, hydroxy, C₁-C₃ alkoxy and C₁-C₃ alkyl; and

R₁₄ is chosen from null, lower cycloalkyl, lower heterocycloalkyl,phenyl, and lower heteroaryl, any of which may be optionallysubstituted.

When, for example, Y₁ is —(CR_(6a)R_(6b))_(m)—Z₁—(CR_(7a)R_(7b))_(n)—,and m and n are both 0, and Z₁ is a bond, then Y₁ collapses to a directbond linking the parent ring system with R₁. This applies to all similarconstructions used herein, including Y₂ and Y₃. Also, when for exampleY₁ is —(CR_(6a)R_(6b))_(m)—Z₁—(CR_(7a)R_(7b))_(n)—, the rightmostportion of Y₁ attaches to the parent molecule.

In certain embodiments, Y₁, Y₂, Y₃, and Y₄ are no more than 6 atoms inlength.

In certain embodiments, R₄ is chosen from hydrogen, (O), and (S).

In certain embodiments, R₄ is (O), the second bond linking R₄ and thefused bicyclic core is present, and the second bond in the five-memberedportion of the fused bicyclic core is absent.

In certain embodiments, R₄ is hydrogen, the second bond linking R₄ andthe fused bicyclic core is absent, and the second bond in thefive-membered portion of the fused bicyclic core is present.

In certain embodiments,

X₁ is CH; and

X₂ is C.

In certain embodiments,

X₁ is N; and

X₂ is N.

In certain embodiments,

X₁ is CH; and

X₂ is N.

In certain embodiments,

X₁ is N; and

X₂ is C.

In certain embodiments,

m and n are both 0;

Z₁ is a bond; and

R₁ and R₅ are both hydrogen.

In certain embodiments,

p and r are each independently an integer from 0 to 3;

q and s are each 0; and

Z₂ and Z₃ are each independently chosen from a bond and O.

In certain embodiments, R_(6a), R_(6b), R_(7a), R_(7b), R_(8a), R_(8b),R_(9a), R_(9b), R_(10a), R_(10b), R_(11a), R_(11b), and R₁₂ are allhydrogen.

In certain embodiments, compounds have structural Formula II

wherein:

dashed lines indicate that a second bond may alternatively be present orabsent;

X₁ is chosen from CH and N;

X₂ is chosen from C and N;

Y₁, Y₂, and Y₃ are independently chosen from a bond, lower alkyl, lowercarboxyl, and lower heteroalkyl;

Y₄ is chosen from —(CH₂)_(m), C(O), —(CH₂)_(m)O—, and —(CH₂)_(m)N—;

m is an integer from 0 to 2;

R₁, R₂, and R₃ are independently chosen from lower alkyl, lower alkenyl,lower alkynyl, lower haloalkyl, lower cycloalkyl, heterocycloalkyl,aryl, heteroaryl, acyl, amido, amino, alkoxy, hydroxy, cyano, and nitro,any of which may be optionally substituted; or R₁ and R₂ may eachadditionally be heteroalkyl, and may be joined together such that R₁ andR₂ together form an alkylene, alkenylene, or heteroalkyl bridgecomprising from 3 to 5 atoms, which may be optionally substituted;

R₄ is chosen from hydrogen, (O), and (S);

R₅ is chosen from hydrogen, hydroxy, cyano, lower alkyl, lowercycloalkyl, and lower alkoxy, any of which may be optionallysubstituted;

R₁₃ is chosen from hydrogen, halogen, hydroxy, cyano, nitro, loweralkyl, lower cycloalkyl, lower cycloalkylalkyl, and lower alkoxy, any ofwhich may be optionally substituted; and additionally, R₁₃ and R₃ may bejoined together to form a lower spiro-cycloalkyl or spiro-phenylcomprising from 3 to 6 atoms, which may be optionally substituted; and

R₁₄ is chosen from null, lower cycloalkyl, lower heterocycloalkyl,phenyl, and lower heteroaryl, any of which may be optionallysubstituted.

In certain embodiments, compounds have structural Formula III

wherein:

dashed lines indicate that a second bond may alternatively be present orabsent;

X₁ and X₂ are independently chosen from CH and N;

Y₃ is chosen from a bond, lower alkyl, lower carboxyl, and lowerheteroalkyl;

Y₄ is chosen from C(O), —(CH₂)_(m)—, —(CH₂)_(m)O—, and —(CH₂)_(m)N—;

m is an integer from 0 to 1;

R₂ and R₃ are independently chosen from lower cycloalkyl,heterocycloalkyl, aryl, heteroaryl, any of which may be optionallysubstituted;

R₄ is chosen from hydrogen, (O), and (S);

R₁₃ is chosen from hydrogen, lower alkyl, lower cycloalkyl, lowercycloalkylalkyl, and lower alkoxy, any of which may be optionallysubstituted;

R₁₄ is chosen from null, lower cycloalkyl, lower heterocycloalkyl,phenyl, and lower heteroaryl, any of which may be optionallysubstituted.

In certain embodiments, compounds have a structural Formula chosen fromFormula IV and Formula V:

wherein:

X₁ and X₂ are independently chosen from CH and N;

Y₃ is chosen from a bond, lower alkyl, lower carboxyl, and lowerheteroalkyl;

Y₄ is chosen from C(O), —(CH₂)_(m)—, —(CH₂)_(m)O—, and —(CH₂)_(m)N—;

m is an integer from 0 to 1;

R₂ and R₃ are independently chosen from lower cycloalkyl, lowerheterocycloalkyl, lower aryl, and lower heteroaryl, any of which may beoptionally substituted; and

R₁₄ is chosen from null, lower cycloalkyl, lower heterocycloalkyl,phenyl, and lower heteroaryl, any of which may be optionallysubstituted.

In certain embodiments, compounds have a structural Formula chosen fromFormula VI, Formula VII, Formula VIII, and Formula IX:

wherein

Y₃ is chosen from a bond, lower alkyl, lower carboxyl, and lowerheteroalkyl;

Y₄ is chosen from C(O), —(CH₂)_(m)—, —(CH₂)_(m)O—, and —(CH₂)_(m)N—;

m is an integer from 0 to 1;

R₂ is chosen from phenyl, 6-membered monocyclic heteroaryl, and5/6-fused bicyclic heteroaryl, any of which may be optionallysubstituted;

R₃ is chosen from lower cycloalkyl, phenyl, and lower heteroaryl, any ofwhich may be optionally substituted;

R₁₄ is chosen from null, lower cycloalkyl, lower heterocycloalkyl,phenyl, and lower heteroaryl, any of which may be optionallysubstituted.

In certain embodiments, R₂ and R₃ are each independently chosen fromlower cycloalkyl, lower aryl, and monocyclic or bicyclic heteroaryl, anyof which may be optionally substituted.

In certain embodiments, R₂ is substituted with one or more substituentschosen from halogen, hydroxy, lower amino, C₁-C₃ alkoxy and C₁-C₃ alkyl.

In further embodiments, R₂ is chosen from phenyl and lower heteroaryl,any of which may be optionally substituted.

In further embodiments, R₂ is chosen from phenyl, 6-membered monocyclicheteroaryl, and 5/6-fused bicyclic heteroaryl, any of which may beoptionally substituted.

In further embodiments, R₂ is chosen from phenyl, pyridinyl,pyrimidinyl, and indolyl, any of which may be optionally substituted.

In further embodiments, R₂ is substituted with one or more substituentschosen from fluorine, hydroxy, NH₂, NH(CH₃), N(CH₃)₂, methoxy, andmethyl.

In further embodiments, R₂ is optionally substituted phenyl.

In further embodiments, R₂ is chosen from

wherein

u is an integer from 0 to 3;

Y₄ is chosen from C(O), —(CH₂)_(m)—, —(CH₂)_(m)O—, and —(CH₂)_(m)N—;

m is an integer from 0 to 1;

R₁₄ is chosen from null, lower cycloalkyl, lower heterocycloalkyl,phenyl, and lower heteroaryl, any of which may be optionallysubstituted; and

each R₁₅ is independently chosen from halogen, hydroxy, C₁-C₄ alkyl,C₂-C₄ alkenyl, C₂-C₄ alkynyl, lower amino, lower amido, lowersulfonamido, and lower sulfonyl.

In certain embodiments, R₁₄ is chosen from piperazinyl, morpholinyl,pyrrolyl, and N(CH₃)₂.

In certain embodiments, each R₁₅ is independently chosen from R₁₅ isindependently chosen from fluorine, hydroxy, NH₂, NH(CH₃), N(CH₃)₂,NS(O)₂CH₃, methoxy, and methyl.

In certain embodiments,

Y₄ is —(CH₂)_(m)—;

m is 0;

R₁₄ is null;

u is an integer from 0 to 3; and

R₁₅ is independently chosen from R₁₅ is independently chosen fromfluorine, hydroxy, NH₂, NH(CH₃), N(CH₃)₂, NS(O)₂CH₃, methoxy, andmethyl.

In certain embodiments, Y₄ is chosen from C(O), O, N, and —CH₂—.

In certain embodiments, Y₄ is —CH₂—.

In certain embodiments, Y₃ is chosen from a bond and lower alkyl.

In certain embodiments, Y₃ is chosen from a bond and methyl.

In certain embodiments, Y₃ is a bond.

In certain embodiments, R₃ is chosen from lower cycloalkyl, lower aryl,and monocyclic or bicyclic heteroaryl, any of which may be optionallysubstituted.

In certain embodiments, R₃ is substituted with one or more substituentschosen from halogen, hydroxy, lower amino, lower amido, lowerphenylamido, lower phenylalkylamido, lower heterocycloalkyl,lowerheterocycloalkyl, loweralkylheterocycloalkyl, C₁-C₃ alkoxy andC₁-C₃ alkyl.

In certain embodiments, R₃ is chosen from benzothiazolyl,pyrrolopyridinyl, indanyl, cyclopropyl, cyclopentyl, phenyl, pyridinyl,pyrimidinyl, and indolyl, any of which may be optionally substituted.

In certain embodiments, R₃ is substituted with one or more substituentschosen from fluorine, chlorine, hydroxy, NH₂, NH(CH₃), N(CH₃)₂, C(O)NH₂,C(O)NHCH₃, morpholino, piperazinyl, methylpiperazinyl, acetamido,methylacetamido, methylpropionamido, phenylacetamidomethylene,benzamidomethylene, phenylpropanamidomethylene, methoxy and methyl.

In certain embodiments are provided a compound of structural Formula III

or a salt thereof, wherein:

-   -   dashed lines indicate that a second bond may alternatively be        present or absent;    -   X₁ and X₂ are independently chosen from CH and N;    -   Y₃ is a bond;    -   Y₄ is chosen from C(O), CH₂, CHF, and CF₂;    -   R₂ is chosen from phenyl and 6-membered monocyclic heteroaryl,        either of which may be optionally substituted;    -   R₃ is optionally substituted bicyclic heteroaryl;

R₄ is chosen from hydrogen, (O), and (S);

R₁₄ is optionally substituted monocyclic heterocycloalkyl.

In certain embodiments, R₃ is an optionally substituted 5/6-fusedbicyclic heteroaryl.

In certain embodiments, wherein Y₄ is CH₂.

In certain embodiments, R₁₄ is optionally substituted piperazinyl.

In certain embodiments, R₂ is chosen from hydrogen, halo, hydroxy, C₁-C₄alkyl, C₃-C₁₀ cycloalkyl, C₁-C₄ alkyloxy, C₃-C₁₀ cycloalkyloxy, aryl,cyano or nitro.

In certain embodiments, R₁ and R₂ together form a butadienylene bridge.

In certain embodiments,

m and n are both 0;

Z₁ is a bond;

R₁, R₅, and R₄ are hydrogen; and

R₂ and R₃ are each independently chosen from aryl and heteroaryl, eitherof which may be optionally substituted.

In certain embodiments,

m and n are both 0;

Z₁ is a bond;

R₁, R₅, and R₄ are hydrogen;

R₂ is selected from the group consisting of aryl and heteroaryl, eitherof which may be optionally substituted; and

R₃ is chosen from 5-substituted-1H-indole, 5-substitutedpyridine-2-amine, and 5-substituted pyrimidine-2-amine.

In certain embodiments,

m is 0 or 1

n is 0;

Z₁ is a bond;

R₁, R₅, and R₄ are hydrogen; and

R₁ is chosen from 5-substituted-1H-indole, 5-substitutedpyridine-2-amine, and 5-substituted pyrimidine-2-amine; and

R₂ is chosen from 5-substituted-1,2,3-trimethoxybenzene,4-substituted-1,2-dimethoxyphenyl, 5-substituted pyridine-2-amine, and5-substituted pyrimidine-2-amine

In certain embodiments,

R₁, R₅, and R₄ are hydrogen; and

R₂ and R₃ are each independently chosen from aryl and heteroaryl, eitherof which may be optionally substituted.

In certain embodiments of Formula I,

m and n are both 0;

Z₁ is a bond;

R₁, R₅, and R₄ are hydrogen,

R₂ is chosen from aryl and heteroaryl, either of which may be optionallysubstituted; and

R₃ is chosen from 5-substituted-1H-indole, 5-substitutedpyridine-2-amine, and 5-substituted pyrimidine-2-amine, any of which maybe optionally substituted.

In certain embodiments of Formula I,

m and n are both 0;

Z₁ is a bond;

R₁, R₅, and R₄ represent hydrogen,

R₃ is chosen from 5-substituted-1H-indole, 5-substitutedpyridine-2-amine, and 5-substituted pyrimidine-2-amine; and

R₂ is chosen from 5-substituted-1,2,3-trimethoxybenzene,4-substituted-1,2-dimethoxybenzene, 5-substituted pyridine-2-amine, and5-substituted pyrimidine-2-amine

In certain embodiments,

R₄ is (O), the second bond linking R₄ and the fused bicyclic core ispresent, and the second bond in the five-membered portion of the fusedbicyclic core is absent;

m and n are both 0;

Z₁ is a bond;

R₁ and R₅ are each hydrogen; and

R₂ and R₃ are each independently chosen from aryl and heteroaryl, eitherof which may be optionally substituted.

In certain embodiments,

R₄ is (O), the second bond linking R₄ and the fused bicyclic core ispresent, and the second bond in the five-membered portion of the fusedbicyclic core is absent;

m and n are both 0;

Z₁ is a bond;

R₁ and R₅ are each hydrogen; and

R₂ is chosen from aryl and heteroaryl, either of which may be optionallysubstituted; and

R₃ is chosen from 5-substituted-1H-indole, 5-substitutedpyridine-2-amine, and 5-substituted pyrimidine-2-amine.

In certain embodiments,

R₄ is (O), the second bond linking R₄ and the fused bicyclic core ispresent, and the second bond in the five-membered portion of the fusedbicyclic core is absent;

m and n are both 0;

Z₁ is a bond;

R₁ and R₅ are each hydrogen;

R₃ is chosen from 5-substituted-1H-indole, 5-substitutedpyridine-2-amine, or 5-substituted pyrimidine-2-amine; and

R₂ is chosen from 5-substituted-1,2,3-trimethoxybenzene,4-substituted-1,2-dimethoxybenzene, 5-substituted pyridine-2-amine, and5-substituted pyrimidine-2-amine

In certain embodiments, optionally substituted groups are substitutedwith one or more substituent chosen from halogen, hydroxy, C₁-C₃ alkoxyand C₁-C₃ alkyl.

In certain embodiments, R₄ is mono- or poly-substituted with fluorine.

In certain embodiments, R₅ is mono- or poly-substituted with fluorine.

In certain embodiments is provided a compound chosen from Examples 1 to167.

Also provided herein is a compound as disclosed herein for use as amedicament.

Also provided herein is a compound as disclosed herein for use as amedicament for the treatment of an MLK-mediated disease.

Also provided herein is the use of a compound as disclosed herein in themanufacture of a medicament for the treatment of an MLK-mediateddisease.

Also provided herein is a pharmaceutical composition comprising acompound of Formula I together with a pharmaceutically acceptablecarrier.

Also provided is a pharmaceutical composition comprising a compoundchosen from Examples 1 to 167.

Also provided herein is a method of inhibition of MLK comprisingcontacting MLK with a compound of Formula I.

In certain embodiments, said MLK is MLK3.

In certain embodiments, said inhibition is selective over other kinases.

Also provided herein is a method of treatment of a MLK-mediated diseasecomprising the administration of a therapeutically effective amount of acompound of Formula Ito a patient in need thereof.

In certain embodiments, said disease is an inflammatory disease or ametabolic disease.

In certain embodiments, said disease is chosen from diabetes mellitus,hyperglycemia, retinopathy, nephropathy, neuropathy, ulcers, micro- andmacroangiopathies, gout and diabetic foot disease, insulin resistance,metabolic syndrome, hyperinsulinemia, hypertension, hyperuricemia,obesity, edema, dyslipidemia, chronic heart failure, atherosclerosis,peripheral inflammation, and HIV dementia.

Also provided herein is a method of treatment of a MLK-mediated diseasecomprising the administration of:

a) a therapeutically effective amount of a compound of Formula I; and

b) another therapeutic agent.

In certain embodiments, said disorder a psychological disorder.

In certain embodiments, said disease is chosen from depression, bipolardisorder, and post-traumatic stress disorder (PTSD).

In certain embodiments, said disorder is a traumatic brain injury.

In certain embodiments, said traumatic brain injury is stroke.

In certain embodiments, said disorder is chosen from Alzheimer's Disease(AD), Parkinson's Disease, HIV dementia and HIV associatedneurocognitive disorder (HAND).

In certain embodiments, said disorder is a neurologic disorder ofhearing or vision.

In certain embodiments, said disorder is chosen from ototoxicity,hearing loss, acute injury to the inner ear, acoustic trauma, and injuryresulting from blast noise.

In certain embodiments the methods of treatment disclosed hereinadditionally comprise the administration of a second therapeutic agent,as part of a therapeutic regimen. The compounds may be delivered in thesame dosage form or separately, and further may be taken concurrently orone subsequent to the other.

In certain embodiments, said second therapeutic agent is a selectiveserotonin reuptake inhibitor (SSRI).

In certain embodiments, said second therapeutic agent is CEP1347.

Also provided herein is a method of treatment of a MLK-mediated diseasecomprising the administration of:

a) a therapeutically effective amount of a an MKL inhibitor; and

b) another therapeutic agent.

In certain embodiments, said second therapeutic agent is a selectiveserotonin reuptake inhibitor (SSRI).

In certain embodiments, said second therapeutic agent is CEP1347.

Also provided herein is a method of achieving an effect in a patientcomprising the administration of a therapeutically effective amount of acompound as disclosed herein to a patient, wherein the effect is chosenfrom:

increased survival of cells of the nervous system, cochlear cells,vestibular cells or retinal cells;

increased survival of heart cells;

promotion of neurogenesis;

promotion of synaptogenesis;

prevention or reduction of neuronal damage;

restoration or improvement of neuronal function;

suppression of neuroinflammation or peripheral inflammation;

suppression of activation of immune cells;

suppression of proliferation of hepatocytes following injury; and

suppression of proliferation of cancer cells.

In certain embodiments, the effect is chosen from:

increased survival of heart cells;

suppression of neuroinflammation or peripheral inflammation;

suppression of activation of immune cells;

suppression of proliferation of hepatocytes following injury; and

suppression of proliferation of cancer cells.

In certain embodiments, said immune cells are chosen from monocytes,macrophages and microglia.

In certain embodiments, the effect is chosen from:

increased survival of cells of the nervous system, cochlear cells,vestibular cells or retinal cells;

increased survival of heart cells;

promotion of neurogenesis;

promotion of synaptogenesis;

prevention or reduction of neuronal damage;

restoration or improvement of neuronal function;

suppression of neuroinflammation or peripheral inflammation;

suppression of activation of immune cells;

suppression of proliferation of hepatocytes following injury; and

suppression of proliferation of cancer cells.

In certain embodiments, said immune cells are chosen from monocytes,macrophages and microglia.

In certain embodiments, the effect is chosen from:

increased survival of cells of the nervous system, cochlear cells,vestibular cells or retinal cells;

promotion of neurogenesis;

promotion of synaptogenesis;

prevention or reduction of neuronal damage; and

restoration or improvement of neuronal function.

As used herein, the terms below have the meanings indicated.

When ranges of values are disclosed, and the notation “from n₁ . . . ton₂” is used, where n₁ and n₂ are the numbers, then unless otherwisespecified, this notation is intended to include the numbers themselvesand the range between them. This range may be integral or continuousbetween and including the end values. By way of example, the range “from2 to 6 carbons” is intended to include two, three, four, five, and sixcarbons, since carbons come in integer units. Compare, by way ofexample, the range “from 1 to 3 μM (micromolar),” which is intended toinclude 1 μM, 3 μM, and everything in between to any number ofsignificant figures (e.g., 1.255 μM, 2.1 μM, 2.9999 μM, etc.).

The term “about,” as used herein, is intended to qualify the numericalvalues which it modifies, denoting such a value as variable within amargin of error. When no particular margin of error, such as a standarddeviation to a mean value given in a chart or table of data, is recited,the term “about” should be understood to mean that range which wouldencompass the recited value and the range which would be included byrounding up or down to that figure as well, taking into accountsignificant figures.

The term “acyl,” as used herein, alone or in combination, refers to acarbonyl attached to an alkenyl, alkyl, aryl, cycloalkyl, heteroaryl,heterocycle, or any other moiety were the atom attached to the carbonylis carbon. An “acetyl” group refers to a —C(O)CH₃ group. An“alkylcarbonyl” or “alkanoyl” group refers to an alkyl group attached tothe parent molecular moiety through a carbonyl group. Examples of suchgroups include methylcarbonyl and ethylcarbonyl. Examples of acyl groupsinclude formyl, alkanoyl and aroyl.

The term “alkenyl,” as used herein, alone or in combination, refers to astraight-chain or branched-chain hydrocarbon radical having one or moredouble bonds and containing from 2 to 20 carbon atoms. In certainembodiments, said alkenyl will comprise from 2 to 6 carbon atoms. Theterm “alkenylene” refers to a carbon-carbon double bond system attachedat two or more positions such as ethenylene [R—CH═CH—), (—C::C—)].Examples of suitable alkenyl radicals include ethenyl, propenyl,2-propenyl, 2-methylpropenyl, butenyl, isobutenyl, 1,4-butadienyl,isoprenyl, vinyl, and the like. Unless otherwise specified, the term“alkenyl” may include “alkenylene” groups.

The term “alkoxy,” as used herein, alone or in combination, refers to analkyl ether radical, wherein the term alkyl is as defined below.Examples of suitable alkyl ether radicals include methoxy, ethoxy,n-propoxy, isopropoxy, n-butoxy, iso-butoxy, sec-butoxy, tert-butoxy,and the like.

The term “alkyl,” as used herein, alone or in combination, refers to astraight-chain or branched-chain alkyl radical containing from 1 to 20carbon atoms. In certain embodiments, said alkyl will comprise from 1 to10 carbon atoms. In further embodiments, said alkyl will comprise from 1to 6 carbon atoms. Alkyl groups may be optionally substituted as definedherein. Examples of alkyl radicals include methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl,hexyl, octyl, noyl and the like. The term “alkylene,” as used herein,alone or in combination, refers to a saturated aliphatic group derivedfrom a straight or branched chain saturated hydrocarbon attached at twoor more positions, such as methylene (—CH₂—). Unless otherwisespecified, the term “alkyl” may include “alkylene” groups.

The term “alkylamino,” as used herein, alone or in combination, refersto an alkyl group attached to the parent molecular moiety through anamino group. Suitable alkylamino groups may be mono- or dialkylated,forming groups such as, for example, N-methylamino, N-ethylamino,N,N-dimethylamino, N,N-ethylmethylamino and the like.

The term “alkylidene,” as used herein, alone or in combination, refersto an alkenyl group in which one carbon atom of the carbon-carbon doublebond belongs to the moiety to which the alkenyl group is attached.

The term “alkylthio,” as used herein, alone or in combination, refers toan alkyl thioether (R—S—) radical wherein the term alkyl is as definedabove and wherein the sulfur may be singly or doubly oxidized. Examplesof suitable alkyl thioether radicals include methylthio, ethylthio,n-propylthio, isopropylthio, n-butylthio, iso-butylthio, sec-butylthio,tert-butylthio, methanesulfonyl, ethanesulfinyl, and the like.

The term “alkynyl,” as used herein, alone or in combination, refers to astraight-chain or branched chain hydrocarbon radical having one or moretriple bonds and containing from 2 to 20 carbon atoms. In certainembodiments, said alkynyl comprises from 2 to 6 carbon atoms. In furtherembodiments, said alkynyl comprises from 2 to 4 carbon atoms. The term“alkynylene” refers to a carbon-carbon triple bond attached at twopositions such as ethynylene (—C:::C—, —CC—). Examples of alkynylradicals include ethynyl, propynyl, hydroxypropynyl, butyn-1-yl,Butyn-2-yl, pentyn-1-yl, 3-methylbutyn-1-yl, hexyn-2-yl, and the like.Unless otherwise specified, the term “alkynyl” may include “alkynylene”groups.

The terms “amido” and “carbamoyl,” as used herein, alone or incombination, refer to an amino group as described below attached to theparent molecular moiety through a carbonyl group, or vice versa. Theterm “C-amido” as used herein, alone or in combination, refers to a—C(O)N(RR′) group with R and R′ as defined herein or as defined by thespecifically enumerated “R” groups designated. The term “N-amido” asused herein, alone or in combination, refers to a RC(O)N(R′)— group,with R and R′ as defined herein or as defined by the specificallyenumerated “R” groups designated. The term “acylamino” as used herein,alone or in combination, embraces an acyl group attached to the parentmoiety through an amino group. An example of an “acylamino” group isacetylamino (CH₃C(O)NH—).

The term “amino,” as used herein, alone or in combination, refers to—NRR′, wherein R and R′ are independently chosen from hydrogen, alkyl,acyl, heteroalkyl, aryl, cycloalkyl, heteroaryl, and heterocycloalkyl,any of which may themselves be optionally substituted. Additionally, Rand R′ may combine to form heterocycloalkyl, either of which may beoptionally substituted.

The term “aryl,” as used herein, alone or in combination, means acarbocyclic aromatic system containing one, two or three rings whereinsuch polycyclic ring systems are fused together. The term “aryl”embraces aromatic groups such as phenyl, naphthyl, anthracenyl, andphenanthryl.

The term “arylalkenyl” or “aralkenyl,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkenyl group.

The term “arylalkoxy” or “aralkoxy,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkoxy group.

The term “arylalkyl” or “aralkyl,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkyl group.

The term “arylalkynyl” or “aralkynyl,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkynyl group.

The term “arylalkanoyl” or “aralkanoyl” or “aroyl,” as used herein,alone or in combination, refers to an acyl radical derived from anaryl-substituted alkanecarboxylic acid such as benzoyl, naphthoyl,phenylacetyl, 3-phenylpropionyl (hydrocinnamoyl), 4-phenylbutyryl,(2-naphthyl)acetyl, 4-chlorohydrocinnamoyl, and the like.

The term aryloxy as used herein, alone or in combination, refers to anaryl group attached to the parent molecular moiety through an oxy.

The terms “benzo” and “benz,” as used herein, alone or in combination,refer to the divalent radical C₆H₄═ derived from benzene. Examplesinclude benzothiophene and benzimidazole.

The term “carbamate,” as used herein, alone or in combination, refers toan ester of carbamic acid (—NRC(O)O—) which may be attached to theparent molecular moiety from either the nitrogen or acid end, and whichmay be optionally substituted as defined herein. The term “O-carbamyl”as used herein, alone or in combination, refers to a —OC(O)NRR′ group;and the term “N-carbamyl” as used herein, alone or in combination,refers to a ROC(O)NR′— group. R and R′ are as defined herein, or asdefined by the specifically enumerated “R” groups designated.

The term “carbonyl,” as used herein, when alone includes formyl[—C(O)H]and in combination is a —C(O)— group.

The term “carboxyl” or “carboxy,” as used herein, refers to —C(O)OH orthe corresponding “carboxylate” anion, such as is in a carboxylic acidsalt. An “O-carboxy” group refers to a RC(O)O— group, where R is asdefined herein. A “C-carboxy” group refers to a —C(O)OR groups where Ris as defined herein.

The term “cyano,” as used herein, alone or in combination, refers to—CN.

The term “cycloalkyl,” or, alternatively, “carbocycle,” as used herein,alone or in combination, refers to a saturated or partially saturatedmonocyclic, bicyclic or tricyclic alkyl group wherein each cyclic moietycontains from 3 to 12 carbon atom ring members and which may optionallybe a benzo fused ring system which is optionally substituted as definedherein. In certain embodiments, said cycloalkyl will comprise from 5 to7 carbon atoms. Examples of such cycloalkyl groups include cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, tetrahydronaphthyl,indanyl, octahydronaphthyl, 2,3-dihydro-1H-indenyl, adamantyl and thelike. “Bicyclic” and “tricyclic” as used herein are intended to includeboth fused ring systems, such as decahydronaphthalene,octahydronaphthalene as well as the multicyclic (multicentered)saturated or partially unsaturated type. The latter type of isomer isexemplified in general by, bicyclo[1,1,1]pentane, camphor, adamantane,and bicyclo[3,2,1]octane.

The term “ester,” as used herein, alone or in combination, refers to acarboxy group bridging two moieties linked at carbon atoms.

The term “ether,” as used herein, alone or in combination, refers to anoxy group bridging two moieties linked at carbon atoms.

The term “halo,” or “halogen,” as used herein, alone or in combination,refers to fluorine, chlorine, bromine, or iodine.

The term “haloalkoxy,” as used herein, alone or in combination, refersto a haloalkyl group attached to the parent molecular moiety through anoxygen atom. Haloalkoxy includes perhaloalkoxy. The term “perhaloalkoxy”refers to an alkoxy group where all of the hydrogen atoms are replacedby halogen atoms. An example of perhaloalkoxy is perfluoromethoxy.

The term “haloalkyl,” as used herein, alone or in combination, refers toan alkyl radical having the meaning as defined above wherein one or morehydrogens are replaced with a halogen. Specifically embraced aremonohaloalkyl, dihaloalkyl, polyhaloalkyl, and perhaloalkyl radicals. Amonohaloalkyl radical, for one example, may have an iodo, bromo, chloroor fluoro atom within the radical. Dihalo and polyhaloalkyl radicals mayhave two or more of the same halo atoms or a combination of differenthalo radicals. Examples of haloalkyl radicals include fluoromethyl,difluoromethyl, trifluoromethyl, chloromethyl, dichloromethyl,trichloromethyl, pentafluoroethyl, heptafluoropropyl,difluorochloromethyl, dichlorofluoromethyl, difluoroethyl,difluoropropyl, dichloroethyl and dichloropropyl. “Haloalkylene” refersto a haloalkyl group attached at two or more positions. Examples ofhaloalkyl radicals include fluoromethyl, difluoromethyl,trifluoromethyl, chloromethyl, dichloromethyl, trichloromethyl,pentafluoroethyl, heptafluoropropyl, difluorochloromethyl,dichlorofluoromethyl, difluoroethyl, difluoropropyl, dichloroethyl anddichloropropyl. “Haloalkylene” refers to a haloalkyl group attached attwo or more positions. Examples include fluoromethylene (—CFH—),difluoromethylene (—CF₂—), chloromethylene (—CHCl—) and the like. Theterm “perhaloalkyl” as used herein, alone or in combination, refers toan alkyl group where all of the hydrogen atoms are replaced by halogenatoms. Examples include perfluoromethyl.

The term “heteroalkyl,” as used herein, alone or in combination, refersto a stable straight or branched chain, or cyclic hydrocarbon radical,or combinations thereof, fully saturated or containing from 1 to 3degrees of unsaturation, consisting of the stated number of carbon atomsand from one to three heteroatoms chosen from O, N, and S, and whereinthe nitrogen and sulfur atoms may optionally be oxidized and thenitrogen heteroatom may optionally be quaternized. The heteroatom(s) O,N and S may be placed at any interior position of the heteroalkyl group.Up to two heteroatoms may be consecutive, such as, for example,—CH₂—NH—OCH₃.

The term “heteroaryl,” as used herein, alone or in combination, refersto a 3 to 15 membered unsaturated heteromonocyclic ring, or a fusedmonocyclic, bicyclic, or tricyclic ring system in which at least one ofthe fused rings is aromatic, which contains at least one atom chosenfrom O, S, and N. Additionally, a heteroaryl may contain one or twoC(O), S(O), or S(O)₂ groups as ring members. In certain embodiments,said heteroaryl will comprise from 5 to 10 atoms. In certainembodiments, said heteroaryl will comprise from 5 to 7 atoms. In certainembodiments, said heteroaryl will comprise from 1 to 4 heteroatoms asring members. In further embodiments, said heteroaryl will comprise from1 to 2 heteroatoms as ring members. The term also embraces fusedpolycyclic groups wherein heterocyclic rings are fused with aryl rings,wherein heteroaryl rings are fused with other heteroaryl rings, whereinheteroaryl rings are fused with heterocycloalkyl rings, or whereinheteroaryl rings are fused with cycloalkyl rings. Examples of heteroarylgroups include pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridyl,pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, triazinyl, triazolyl,tetrazolyl, pyranyl, furyl, thienyl, oxazolyl, isoxazolyl, oxadiazolyl,thiazolyl, thiadiazolyl, isothiazolyl, indolyl, isoindolyl, indolizinyl,benzimidazolyl, quinolyl, isoquinolyl, quinoxalinyl, quinazolinyl,indazolyl, benzotriazolyl, benzodioxolyl, benzopyranyl, benzoxazolyl,benzoxadiazolyl, benzothiazolyl, benzothiadiazolyl, benzofuryl,benzothienyl, chromonyl, coumarinyl, benzopyranyl, tetrahydroquinolinyl,tetrazolopyridazinyl, tetrahydroisoquinolinyl, thienopyridinyl,furopyridinyl, pyrrolopyridinyl and the like. Exemplary tricyclicheterocyclic groups include carbazolyl, benzidolyl, phenanthrolinyl,dibenzofuranyl, acridinyl, phenanthridinyl, xanthenyl and the like.

The terms “heterocycloalkyl” and, interchangeably, “heterocycle,” asused herein, alone or in combination, each refer to a saturated,partially unsaturated, or fully unsaturated monocyclic, bicyclic, ortricyclic heterocyclic group containing at least one heteroatom as aring member, wherein each said heteroatom may be independently chosenfrom N, O, and S. Additionally, a heterocycloalkyl may contain one ortwo C(O), S(O), or S(O)₂ groups as ring members. In certain embodiments,said heterocycloalkyl will comprise from 1 to 4 heteroatoms as ringmembers. In further embodiments, said heterocycloalkyl will comprisefrom 1 to 2 heteroatoms as ring members. In certain embodiments, saidheterocycloalkyl will comprise from 3 to 8 ring members in each ring. Infurther embodiments, said heterocycloalkyl will comprise from 3 to 7ring members in each ring. In yet further embodiments, saidheterocycloalkyl will comprise from 5 to 6 ring members in each ring.“Heterocycloalkyl” and “heterocycle” are intended to include sulfones,sulfoxides, N-oxides of tertiary nitrogen ring members, and carbocyclicfused and benzo fused ring systems; additionally, both terms alsoinclude systems where a heterocycle ring is fused to an aryl group, asdefined herein, or an additional heterocycle group. Examples ofheterocycle groups include aziridinyl, azetidinyl, 1,3-benzodioxolyl,dihydroisoindolyl, dihydroisoquinolinyl, dihydrocinnolinyl,dihydrobenzodioxinyl, dihydro[1,3]oxazolo[4,5-b]pyridinyl,benzothiazolyl, dihydroindolyl, dihy-dropyridinyl, 1,3-dioxanyl,1,4-dioxanyl, 1,3-dioxolanyl, isoindolinyl, morpholinyl, piperazinyl,pyrrolidinyl, tetrahydropyridinyl, piperidinyl, thiomorpholinyl, and thelike. The heterocycle groups may be optionally substituted unlessspecifically prohibited.

The term “hydroxy,” as used herein, alone or in combination, refers to—OH.

The term “lower,” as used herein, alone or in a combination, where nototherwise specifically defined, means containing from 1 to and including6 carbon atoms.

The term “lower alkyl,” as used herein, alone or in a combination, meansC₁-C₆ straight or branched chain alkyl. The term “lower alkenyl” meansC₂-C₆ straight or branched chain alkenyl. The term “lower alkynyl” meansC₂-C₆ straight or branched chain alkynyl.

The term “lower aryl,” as used herein, alone or in combination, meansphenyl or naphthyl, either of which may be optionally substituted asprovided.

The term “lower heteroaryl,” as used herein, alone or in combination,means either 1) monocyclic heteroaryl comprising five or six ringmembers, of which between one and four said members may be heteroatomschosen from O, S, and N, or 2) bicyclic heteroaryl, wherein each of thefused rings comprises five or six ring members, comprising between themone to four heteroatoms chosen from O, S, and N.

The term “lower cycloalkyl,” as used herein, alone or in combination,means a monocyclic cycloalkyl having between three and six ring members.Lower cycloalkyls may be unsaturated. Examples of lower cycloalkylinclude cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

The term “lower heterocycloalkyl,” as used herein, alone or incombination, means a monocyclic heterocycloalkyl having between threeand six ring members, of which between one and four may be heteroatomschosen from O, S, and N. Examples of lower heterocycloalkyls includepyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, piperazinyl,and morpholinyl. Lower heterocycloalkyls may be unsaturated.

The term “lower amino,” as used herein, alone or in combination, refersto —NRR′, wherein R and R′ are independently chosen from hydrogen, loweralkyl, and lower heteroalkyl, any of which may be optionallysubstituted. Additionally, the R and R′ of a lower amino group maycombine to form a five- or six-membered heterocycloalkyl, either ofwhich may be optionally substituted.

The term “nitro,” as used herein, alone or in combination, refers to—NO₂.

The terms “oxy” or “oxa,” as used herein, alone or in combination, referto —O—.

The term “oxo,” as used herein, alone or in combination, refers to ═O.

The term “perhaloalkoxy” refers to an alkoxy group where all of thehydrogen atoms are replaced by halogen atoms.

The term “perhaloalkyl” as used herein, alone or in combination, refersto an alkyl group where all of the hydrogen atoms are replaced byhalogen atoms.

The terms “sulfonate,” “sulfonic acid,” and “sulfonic,” as used herein,alone or in combination, refer the —SO₃H group and its anion as thesulfonic acid is used in salt formation.

The term “N-sulfonamido” refers to a RS(═O)₂NR′— group with R and R′ asdefined herein or as defined by the specifically enumerated “R” groupsdesignated.

The term “S-sulfonamido” refers to a —S(═O)₂NRR′, group, with R and R′as defined herein or as defined by the specifically enumerated “R”groups designated.

The terms “thia” and “thio,” as used herein, alone or in combination,refer to a —S— group or an ether wherein the oxygen is replaced withsulfur. The oxidized derivatives of the thio group, namely sulfinyl andsulfonyl, are included in the definition of thia and thio. The term“sulfanyl,” as used herein, alone or in combination, refers to —S—. Theterm “sulfinyl,” as used herein, alone or in combination, refers to—S(O)—. The term “sulfonyl,” as used herein, alone or in combination,refers to —S(O)₂—.

The term “thiol,” as used herein, alone or in combination, refers to an—SH group.

The term “thiocarbonyl,” as used herein, when alone includes thioformyl—C(S)H and in combination is a —C(S)— group.

Any definition herein may be used in combination with any otherdefinition to describe a composite structural group. By convention, thetrailing element of any such definition is that which attaches to theparent moiety. For example, the composite group alkylamido wouldrepresent an alkyl group attached to the parent molecule through anamido group, and the term alkoxyalkyl would represent an alkoxy groupattached to the parent molecule through an alkyl group.

When a group is defined to be “null,” what is meant is that said groupis absent.

The term “optionally substituted” means the anteceding group may besubstituted or unsubstituted. When substituted, the substituents of an“optionally substituted” group may include, without limitation, one ormore substituents independently selected from the following groups or aparticular designated set of groups, alone or in combination: loweralkyl, lower alkenyl, lower alkynyl, lower alkanoyl, lower heteroalkyl,lower heterocycloalkyl, lower haloalkyl, lower haloalkenyl, lowerhaloalkynyl, lower perhaloalkyl, lower perhaloalkoxy, lower cycloalkyl,phenyl, aryl, aryloxy, lower alkoxy, lower haloalkoxy, oxo, loweracyloxy, carbonyl, carboxyl, lower alkylcarbonyl, lower carboxyester,lower carboxamido, cyano, hydrogen, halogen, hydroxy, amino, loweralkylamino, arylamino, amido, nitro, thiol, lower alkylthio, lowerhaloalkylthio, lower perhaloalkylthio, arylthio, sulfonate, sulfonicacid, trisubstituted silyl, N₃, SH, SCH₃, C(O)CH₃, CO₂CH₃, CO₂H,pyridinyl, thiophene, furanyl, lower carbamate, and lower urea. Twosubstituents may be joined together to form a fused five-, six-, orseven-membered carbocyclic or heterocyclic ring consisting of zero tothree heteroatoms, for example forming methylenedioxy or ethylenedioxy.An optionally substituted group may be unsubstituted (e.g., —CH₂CH₃),fully substituted (e.g., —CF₂CF₃), monosubstituted (e.g., —CH₂CH₂F) orsubstituted at a level anywhere in-between fully substituted andmonosubstituted (e.g., —CH₂CF₃). Where substituents are recited withoutqualification as to substitution, both substituted and unsubstitutedforms are encompassed. Where a substituent is qualified as“substituted,” the substituted form is specifically intended.Additionally, different sets of optional substituents to a particularmoiety may be defined as needed; in these cases, the optionalsubstitution will be as defined, often immediately following the phrase,“optionally substituted with.”

The term R or the term R′, appearing by itself and without a numberdesignation, unless otherwise defined, refers to a moiety chosen fromhydrogen, alkyl, cycloalkyl, heteroalkyl, aryl, heteroaryl andheterocycloalkyl, any of which may be optionally substituted. Such R andR′ groups should be understood to be optionally substituted as definedherein. Whether an R group has a number designation or not, every Rgroup, including R, R′ and R^(n) where n=(1, 2, 3, . . . n), everysubstituent, and every term should be understood to be independent ofevery other in terms of selection from a group. Should any variable,substituent, or term (e.g. aryl, heterocycle, R, etc.) occur more thanone time in a formula or generic structure, its definition at eachoccurrence is independent of the definition at every other occurrence.Those of skill in the art will further recognize that certain groups maybe attached to a parent molecule or may occupy a position in a chain ofelements from either end as written. Thus, by way of example only, anunsymmetrical group such as —C(O)N(R)— may be attached to the parentmoiety at either the carbon or the nitrogen.

Asymmetric centers exist in the compounds disclosed herein. Thesecenters are designated by the symbols “R” or “S,” depending on theconfiguration of substituents around the chiral carbon atom. It shouldbe understood that the invention encompasses all stereochemical isomericforms, including diastereomeric, enantiomeric, and epimeric forms, aswell as d-isomers and 1-isomers, and mixtures thereof. Individualstereoisomers of compounds can be prepared synthetically fromcommercially available starting materials which contain chiral centersor by preparation of mixtures of enantiomeric products followed byseparation such as conversion to a mixture of diastereomers followed byseparation or recrystallization, chromatographic techniques, directseparation of enantiomers on chiral chromatographic columns, or anyother appropriate method known in the art. Compounds can be preparedusing diastereomers, enantiomers or racemic mixtures as startingmaterials. Starting compounds of particular stereochemistry are eithercommercially available or can be made and resolved by techniques knownin the art. Furthermore, diastereomer and enantiomer products can beseparated by chromatography, fractional crystallization or other methodsknown to those of skill in the art. Additionally, the compoundsdisclosed herein may exist as geometric isomers. The present inventionincludes all cis, trans, syn, anti, entgegen (E), and zusammen (Z)isomers as well as the appropriate mixtures thereof. Additionally,compounds may exist as tautomers; all tautomeric isomers are provided bythis invention. Solvates, hydrates, isomorphs, polymorphs are alsoprovided. Additionally, the compounds disclosed herein can exist inunsolvated as well as solvated forms with pharmaceutically acceptablesolvents such as water, ethanol, and the like. In general, the solvatedforms are considered equivalent to the unsolvated forms.

The term “bond” refers to a covalent linkage between two atoms, or twomoieties when the atoms joined by the bond are considered to be part oflarger substructure. A bond may be single, double, or triple unlessotherwise specified. A dashed line between two atoms in a drawing of amolecule indicates that an additional bond may be present or absent atthat position. When, for example, Y₁ is—(CR_(6a)R_(6b))_(m)—Z₁—(CR_(7a)R_(7b))_(n)—, and m and n are both 0,and Z₁ is a bond, then Y₁ collapses to a direct bond linking the parentring system with R₁. This applies to all similar constructions usedherein, including Y₂ and Y₃. Or, for example, when either of R_(6a) andR_(6b) of (CR_(6a)R_(6b))—, are designated to be “a bond,” and m≧1, thenan additional bond forms between a C of (CR_(6a)R_(6b)) and an adjacentatom. When m≧2, then (CR_(6a)R_(6b))_(m) may form an alkene (alkenylene)or alkyne (alkynylene).

As used herein, the terms “treating” and “treatment” refer to delayingthe onset of, retarding or reversing the progress of, or alleviating orpreventing either the disease or condition to which the term applies, orone or more symptoms of such disease or condition.

The term “patient” (and, equivalently, “subject”) means all mammalsincluding humans. Examples of patients include humans, cows, dogs, cats,goats, sheep, pigs, and rabbits. Preferably, the patient is a human.

The term “disease” as used herein is intended to be generallysynonymous, and is used interchangeably with, the terms “disorder,”“syndrome,” and “condition” (as in medical condition), in that allreflect an abnormal condition of the human or animal body or of one ormore of its parts that impairs normal functioning, is typicallymanifested by distinguishing signs and symptoms, and/or causes the humanor animal to have a reduced duration or quality of life.

The term “neuropsychiatric disorder” includes, without limitation,psychological, psychiatric, and neurological disorders.

The term “HIV associated neurocognitive disorder (HAND)” is related to,and is intended to be substantially synonymous with, the terms HIVdementia, AIDS dementia, HIV encephalopathy, and NeuroAIDS.

The term “combination therapy” means the administration of two or moretherapeutic agents to treat a therapeutic condition or disorderdescribed in the present disclosure. Such administration encompassesco-administration of these therapeutic agents in a substantiallysimultaneous manner, such as in a single capsule having a fixed ratio ofactive ingredients or in multiple, separate capsules for each activeingredient. In addition, such administration also encompasses use ofeach type of therapeutic agent in a sequential manner. In either case,the treatment regimen will provide beneficial effects of the drugcombination in treating the conditions or disorders described herein.

As used herein, the term “administering” means oral administration,administration as a suppository, topical contact, intravenous,intraperitoneal, intramuscular, intralesional, intranasal orsubcutaneous administration, or the implantation of a slow-releasedevice, e.g., a mini-osmotic pump, to a subject. Administration is byany route including parenteral, and transmucosal (e.g., oral, nasal,vaginal, rectal, or transdermal). Parenteral administration includes,e.g., intravenous, intramuscular, intra-arteriole, intradermal,subcutaneous, intraperitoneal, intraventricular, and intracranial. Othermodes of delivery include, but are not limited to, the use of liposomalformulations, intravenous infusion, transdermal patches, and the like.

As used herein, the term “prodrug” refers to a precursor compound that,following administration, releases the biologically active compound invivo via some chemical or physiological process (e.g., a prodrug onreaching physiological pH or through enzyme action is converted to thebiologically active compound).

The terms “controlled release,” “sustained release,” “extended release,”and “timed release” are intended to refer interchangeably to anydrug-containing formulation in which release of the drug is notimmediate, i.e., with a “controlled release” formulation, oraladministration does not result in immediate release of the drug into anabsorption pool. The terms are used interchangeably with “nonimmediaterelease” as defined in Remington: The Science and Practice of Pharmacy,2E′ Ed., Gennaro, Ed., Lippencott Williams & Wilkins (2003). Asdiscussed therein, immediate and nonimmediate release can be definedkinetically by reference to the following equation:

The “absorption pool” represents a solution of the drug administered ata particular absorption site, and k_(r), k_(a) and k_(e) are first-orderrate constants for (1) release of the drug from the formulation, (2)absorption, and (3) elimination, respectively. For immediate releasedosage forms, the rate constant for drug release k_(r) is far greaterthan the absorption rate constant k_(a). For controlled releaseformulations, the opposite is true, i.e., k_(r)<<k_(a), such that therate of release of drug from the dosage form is the rate-limiting stepin the delivery of the drug to the target area.

The terms “sustained release” and “extended release” are used in theirconventional sense to refer to a drug formulation that provides forgradual release of a drug over an extended period of time, for example,12 hours or more, and that preferably, although not necessarily, resultsin substantially constant blood levels of a drug over an extended timeperiod.

As used herein, the term “delayed release” refers to a pharmaceuticalpreparation that passes through the stomach intact and dissolves in thesmall intestine.

“MLK3 inhibitor” is used herein to refer to a compound that exhibits anIC₅₀ with respect to MLK3 activity of no more than about 100 μM and moretypically not more than about 50 μM, as measured in the MLK3 (assayname) described generally hereinbelow. “IC₅₀” is that concentration ofinhibitor which reduces the activity and/or expression of an enzyme(e.g., MLK or MLK3) to half-maximal level. Certain compounds disclosedherein have been discovered to exhibit inhibition against MLK3. Incertain embodiments, compounds will exhibit an 1050 with respect to MLK3of no more than about 10 μM; in further embodiments, compounds willexhibit an IC₅₀ with respect to MLK3 of no more than about 5 μM; in yetfurther embodiments, compounds will exhibit an IC₅₀ with respect to MLK3of not more than about 1 μM; in yet further embodiments, compounds willexhibit an IC₅₀ with respect to MLK3 of not more than about 200 nM, asmeasured in the MLK3 assay described herein.

The phrase “therapeutically effective” is intended to qualify the amountof active ingredients used in the treatment of a disease or disorder.This amount will achieve the goal of reducing or eliminating the saiddisease or disorder.

The term “therapeutically acceptable” refers to those compounds (orsalts, prodrugs, tautomers, zwitterionic forms, etc.) which are suitablefor use in contact with the tissues of patients without undue toxicity,irritation, and allergic response, are commensurate with a reasonablebenefit/risk ratio, and are effective for their intended use.

The term “prodrug” refers to a compound that is made more active invivo. Certain compounds disclosed herein may also exist as prodrugs, asdescribed in Hydrolysis in Drug and Prodrug Metabolism: Chemistry,Biochemistry, and Enzymology (Testa, Bernard and Mayer, Joachim M.Wiley-VHCA, Zurich, Switzerland 2003). Prodrugs of the compoundsdescribed herein are structurally modified forms of the compound thatreadily undergo chemical changes under physiological conditions toprovide the compound. Additionally, prodrugs can be converted to thecompound by chemical or biochemical methods in an ex vivo environment.For example, prodrugs can be slowly converted to a compound when placedin a transdermal patch reservoir with a suitable enzyme or chemicalreagent. Prodrugs are often useful because, in some situations, they maybe easier to administer than the compound, or parent drug. They may, forinstance, be bioavailable by oral administration whereas the parent drugis not. The prodrug may also have improved solubility in pharmaceuticalcompositions over the parent drug. A wide variety of prodrug derivativesare known in the art, such as those that rely on hydrolytic cleavage oroxidative activation of the prodrug. An example, without limitation, ofa prodrug would be a compound which is administered as an ester (the“prodrug”), but then is metabolically hydrolyzed to the carboxylic acid,the active entity. Additional examples include peptidyl derivatives of acompound.

Prodrugs of compounds of Formula I are provided herein. Prodrugs ofcompounds provided herein include, but are not limited to, carboxylateesters, carbonate esters, hemi-esters, phosphorus esters, nitro esters,sulfate esters, sulfoxides, amides, carbamates, azo compounds,phosphamides, glycosides, ethers, acetals, and ketals. Prodrug estersand carbonates may be formed, for example, by reacting one or morehydroxyl groups of compounds of Formula I or Formula II with alkyl,alkoxy or aryl substituted acylating reagents using methods known tothose of skill in the art to produce methyl carbonates, acetates,benzoates, pivalates and the like. Illustrative examples of prodrugesters of the compounds provided herein include, but are not limited to,compounds of Formula I having a carboxyl moiety wherein the freehydrogen is replaced by C₁-C₄ alkyl, C₁-C₇ alkanoyloxymethyl,1-((C₁-C₅)alkanoyloxy)ethyl, 1-methyl-1-((C₁-C₅)alkanoyloxy)-ethyl,C₁-C₅ alkoxycarbonyloxymethyl, 1-((C₁-C₅)alkoxycarbonyloxy)ethyl,1-methyl-1-((C₁-C₅)alkoxycarbonyloxy)ethyl,N-((C₁-C₅)alkoxycarbonyl)aminomethyl,1-(N-((C₁-C₅)alkoxycarbonyl)amino)ethyl, 3-phthalidyl,4-crotonolactonyl, gamma-butyrolacton-4-yl,di-N,N-(C₁-C₂)alkylamino(C₂-C₃)alkyl (e.g., beta-dimethylaminoethyl),carbamoyl-(C₁-C₂)alkyl, N,N-di(C₁-C₂)alkylcarbamoyl-(C₁-C₂)alkyl andpiperidino-, pyrrolidino- or morpholino(C₂-C₃)alkyl. Oligopeptidemodifications and biodegradable polymer derivatives (as described, forexample, in Int. J. Pharm. 115, 61-67, 1995) are within the scope of thepresent disclosure. Methods for selecting and preparing suitableprodrugs are provided, for example, in the following: T. Higuchi and V.Stella, “Prodrugs as Novel Delivery Systems,” Vol. 14, ACS SymposiumSeries, 1975; H. Bundgaard, “Design of Prodrugs,” Elsevier, 1985; and“Bioreversible Carriers in Drug Design,” ed. Edward Roche, AmericanPharmaceutical Association and Pergamon Press, 1987.

The compounds disclosed herein can exist as therapeutically acceptablesalts. The present invention includes compounds disclosed herein in theform of salts, including acid addition salts. Suitable salts includethose formed with both organic and inorganic acids. Such acid additionsalts will normally be pharmaceutically acceptable. However, salts ofnon-pharmaceutically acceptable salts may be of utility in thepreparation and purification of the compound in question. Basic additionsalts may also be formed and be pharmaceutically acceptable. For a morecomplete discussion of the preparation and selection of salts, refer toPharmaceutical Salts: Properties, Selection, and Use (Stahl, P.Heinrich. Wiley-VCHA, Zurich, Switzerland, 2002).

The term “therapeutically acceptable salt,” as used herein, representssalts or zwitterionic forms of the compounds disclosed herein which arewater or oil-soluble or dispersible and therapeutically acceptable asdefined herein. The salts can be prepared during the final isolation andpurification of the compounds or separately by reacting the appropriatecompound in the form of the free base with a suitable acid.Representative acid addition salts include acetate, adipate, alginate,L-ascorbate, aspartate, benzoate, benzenesulfonate (besylate),bisulfate, butyrate, camphorate, camphorsulfonate, citrate, digluconate,formate, fumarate, gentisate, glutarate, glycerophosphate, glycolate,hemisulfate, heptanoate, hexanoate, hippurate, hydrochloride,hydrobromide, hydroiodide, 2-hydroxyethansulfonate (isethionate),lactate, maleate, malonate, DL-mandelate, mesitylenesulfonate,methanesulfonate, naphthylenesulfonate, nicotinate,2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate,3-phenylproprionate, phosphonate, picrate, pivalate, propionate,pyroglutamate, succinate, sulfonate, tartrate, L-tartrate,trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate,para-toluenesulfonate (p-tosylate), and undecanoate. Also, basic groupsin the compounds disclosed herein can be quaternized with methyl, ethyl,propyl, and butyl chlorides, bromides, and iodides; dimethyl, diethyl,dibutyl, and diamyl sulfates; decyl, lauryl, myristyl, and sterylchlorides, bromides, and iodides; and benzyl and phenethyl bromides.Examples of acids which can be employed to form therapeuticallyacceptable addition salts include inorganic acids such as hydrochloric,hydrobromic, sulfuric, and phosphoric, and organic acids such as oxalic,maleic, succinic, and citric. Salts can also be formed by coordinationof the compounds with an alkali metal or alkaline earth ion. Hence, thepresent invention contemplates sodium, potassium, magnesium, and calciumsalts of the compounds disclosed herein, and the like.

Basic addition salts can be prepared during the final isolation andpurification of the compounds by reacting a carboxy group with asuitable base such as the hydroxide, carbonate, or bicarbonate of ametal cation or with ammonia or an organic primary, secondary, ortertiary amine. The cations of therapeutically acceptable salts includelithium, sodium, potassium, calcium, magnesium, and aluminum, as well asnontoxic quaternary amine cations such as ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, diethylamine, ethylamine, tributylamine, pyridine,N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine,dicyclohexylamine, procaine, dibenzylamine, N,N-dibenzylphenethylamine,1-ephenamine, and N,N′-dibenzylethylenediamine. Other representativeorganic amines useful for the formation of base addition salts includeethylenediamine, ethanolamine, diethanolamine, piperidine, andpiperazine.

Also provided herein are isotopically-substituted or -labeled compoundsof Formula I, wherein one or more atoms are replaced by one or moreatoms having specific atomic mass or mass numbers. Examples of isotopesthat can be incorporated into compounds disclosed herein include, butare not limited to, isotopes of hydrogen, carbon, nitrogen, oxygen,fluorine, sulfur, and chlorine (such as ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O,¹⁸F, ³⁵S and ³⁶Cl). Isotopically-labeled compounds of Formula I andprodrugs thereof, as well as isotopically-labeled, pharmaceuticallyacceptable salts of compounds of Formula I and prodrugs thereof, areherein disclosed. Isotopically-labeled compounds are useful in assays ofthe tissue distribution of the compounds and their prodrugs andmetabolites; preferred isotopes for such assays include ³H and ¹⁴C. Inaddition, in certain circumstances substitution with heavier isotopes,such as deuterium (²H), can provide increased metabolic stability, whichoffers therapeutic advantages such as increased in vivo half-life orreduced dosage requirements. Isotopically-labeled compounds and prodrugsthereof can generally be prepared according to the methods describedherein by substituting an isotopically-labeled reagent for anon-isotopically labeled reagent.

In other aspects, provided herein are intermediates and processes usefulfor preparing the intermediates below as well as the compounds ofFormula I, and pharmaceutically acceptable salts and prodrugs thereof.

In a similar manner, the present invention provides methods of preparingcompounds of Formula I, that are based on the synthetic protocolsoutlined in Schemes 1 through 21 as well as methods well known bypersons skilled in the art, and the more detailed particular examplespresented below in the experimental section describing the examples. Byfollowing the general preparative methods discussed below, or employingvariations or alternative methods, the compounds can be readily preparedby the use of chemical reactions and procedures known to those of skillin the art. Unless otherwise specified, the variables (e.g., R groups)denoting groups in the general methods described below have the meaningsas hereinbefore defined.

Those of skill in the art will recognize that compounds with eachdescribed functional group are generally prepared using slightvariations of the below-listed general methods. Within the scope of eachmethod, functional groups which are suitable to the reaction conditionsare used. Functional groups which might interfere with certain reactionsare presented in protected forms where necessary, and the removal ofsuch protective groups is completed at appropriate stages by methodswell known to those skilled in the art.

In certain cases compounds can be prepared from other compoundsdisclosed herein by elaboration, transformation, exchange and the likeof the functional groups present. Such elaboration includes, but is notlimited to, hydrolysis, reduction, oxidation, alkylation, acylation,esterification, amidation and dehydration. Such transformations can insome instances require the use of protecting groups by the methodsdisclosed in T. W. Greene and P. G. M. Wuts, Protective Groups inOrganic Synthesis; Wiley: New York, (1999), and incorporated herein byreference. Such methods would be initiated after synthesis of thedesired compound or at another place in the synthetic route that wouldbe readily apparent to one skilled in the art.

In another aspect, provided herein are synthetic intermediates usefulfor preparing the compounds of Formula I, and pharmaceuticallyacceptable salts and prodrugs thereof, according to the generalpreparative methods discussed below and other processes known to thoseof skill in the art.

When the following abbreviations and acronyms are used throughout thedisclosure, they have the following meanings: CDCl₃, chloroform-d;CH₂Cl₂, methylene chloride; CH₃CN, acetonitrile; DIPEA,N,N-diisopropylethylamine; DMAP, 4-dimethylaminopyridine; DMF,N,N-dimethylformamide; DMSO, dimethylsulfoxide; Et, ethyl; Et₃N,triethylamine; EtOAc (or AcOEt), ethyl acetate; EtOH, ethanol; h, hour;HCl, hydrochloric acid; ¹H NMR, proton nuclear magnetic resonance;H₂SO₄, sulfuric acid; HPLC, high performance liquid chromatography;K₂CO₃, potassium carbonate; KOH, potassium hydroxide; LC-MS, liquidchromatography—mass spectroscopy; Me, methyl; MeOH, methanol; min,minute; MS ESI, mass spectroscopy with electrospray ionization; MsOH,methanesulfonic acid; NaH, sodium hydride; NaHCO₃, sodium bicarbonate;NaOH, sodium hydroxide; Na₂SO₄, sodium sulfate; NBS, N-bromosuccinimide;NCS, N-chlorosuccinimide; NH₃, ammonia; NIS, N-iodosuccinimide; Pd/C,palladium on carbon; Pd(PPh₃)₄,tetrakis(triphenylphosphine)palladium(0); Rf, retention factor; TBAF,tetrabutylammonium fluoride; TBAI, tetrabutylammonium iodide; TBDMS,t-butyldimethylsilyl; Tf₂O, trifluoromethanesulfonic anhydride; TFA,trifluoroacetic acid; THF, tetrahydrofuran; TLC, thin layerchromatography; TMS, trimethylsilyl; TMSCN, trimethylsilyl cyanide;TsOH, toluenesulfonic acid.

While it may be possible for compounds to be administered as the rawchemical, it is also possible to present them as a pharmaceuticalformulation. Accordingly, provided herein are pharmaceuticalformulations which comprise one or more of certain compounds disclosedherein, or one or more pharmaceutically acceptable salts, esters,prodrugs, amides, or solvates thereof, together with one or morepharmaceutically acceptable carriers thereof and optionally one or moreother therapeutic ingredients. The carrier(s) must be “acceptable” inthe sense of being compatible with the other ingredients of theformulation and not deleterious to the recipient thereof. Properformulation is dependent upon the route of administration chosen. Any ofthe well-known techniques, carriers, and excipients may be used assuitable and as understood in the art; e.g., in Remington: The Scienceand Practice of Pharmacy, 2E′ Ed., Gennaro, Ed., Lippencott Williams &Wilkins (2003). The pharmaceutical compositions disclosed herein may bemanufactured in any manner known in the art, e.g., by means ofconventional mixing, dissolving, granulating, dragee-making, levigating,emulsifying, encapsulating, entrapping or compression processes.

A compound as provided herein can be incorporated into a variety offormulations for therapeutic administration, including solid,semi-solid, liquid or gaseous forms. The formulations include thosesuitable for oral, parenteral (including subcutaneous, intradermal,intramuscular, intravenous, intraarticular, and intramedullary),intraperitoneal, transmucosal, transdermal, rectal and topical(including dermal, buccal, sublingual and intraocular) administrationalthough the most suitable route may depend upon for example thecondition and disorder of the recipient. The formulations mayconveniently be presented in unit dosage form and may be prepared by anyof the methods well known in the art of pharmacy. Typically, thesemethods include the step of bringing into association a compound or apharmaceutically acceptable salt, ester, amide, prodrug or solvatethereof (“active ingredient”) with the carrier which constitutes one ormore accessory ingredients. In general, the formulations are prepared byuniformly and intimately bringing into association the active ingredientwith liquid carriers or finely divided solid carriers or both and then,if necessary, shaping the product into the desired formulation.

Formulations of the compounds disclosed herein suitable for oraladministration may be presented as discrete units such as capsules,cachets or tablets each containing a predetermined amount of the activeingredient; as a powder or granules; as a solution or a suspension in anaqueous liquid or a non-aqueous liquid; or as an oil-in-water liquidemulsion or a water-in-oil liquid emulsion. The active ingredient mayalso be presented as a bolus, electuary or paste.

Pharmaceutical preparations which can be used orally include tablets,push-fit capsules made of gelatin, as well as soft, sealed capsules madeof gelatin and a plasticizer, such as glycerol or sorbitol. Tablets maybe made by compression or molding, optionally with one or more accessoryingredients. Compressed tablets may be prepared by compressing in asuitable machine the active ingredient in a free-flowing form such as apowder or granules, optionally mixed with binders, inert diluents, orlubricating, surface active or dispersing agents. Molded tablets may bemade by molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent. The tablets may optionally becoated or scored and may be formulated so as to provide slow orcontrolled release of the active ingredient therein. All formulationsfor oral administration should be in dosages suitable for suchadministration. The push-fit capsules can contain the active ingredientsin admixture with filler such as lactose, binders such as starches,and/or lubricants such as talc or magnesium stearate and, optionally,stabilizers. In soft capsules, the active compounds may be dissolved orsuspended in suitable liquids, such as fatty oils, liquid paraffin, orliquid polyethylene glycols. In addition, stabilizers may be added.

Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active compound doses. Also provided are oralformulations in the form of powders and granules containing one or morecompounds disclosed herein.

The compounds may be formulated for parenteral administration byinjection, e.g., by bolus injection or continuous infusion. Formulationsfor injection may be presented in unit dosage form, e.g., in ampoules orin multi-dose containers, with an added preservative. The compositionsmay take such forms as suspensions, solutions or emulsions in oily oraqueous vehicles, and may contain formulatory agents such as suspending,stabilizing and/or dispersing agents. The formulations may be presentedin unit-dose or multi-dose containers, for example sealed ampoules andvials, and may be stored in powder form or in a freeze-dried(lyophilized) condition requiring only the addition of the sterileliquid carrier, for example, saline or sterile pyrogen-free water,immediately prior to use. Extemporaneous injection solutions andsuspensions may be prepared from sterile powders, granules and tabletsof the kind previously described.

Formulations for parenteral administration include aqueous andnon-aqueous (oily) sterile injection solutions of the active compoundswhich may contain antioxidants, buffers, bacteriostats and solutes whichrender the formulation isotonic with the blood of the intendedrecipient; and aqueous and non-aqueous sterile suspensions which mayinclude suspending agents and thickening agents. Suitable lipophilicsolvents or vehicles include fatty oils such as sesame oil, or syntheticfatty acid esters, such as ethyl oleate or triglycerides, or liposomes.Aqueous injection suspensions may contain substances which increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol, or dextran. Optionally, the suspension may also containsuitable stabilizers or agents which increase the solubility of thecompounds to allow for the preparation of highly concentrated solutions.

In addition to the formulations described previously, the compounds mayalso be formulated as a depot preparation. Such long acting formulationsmay be administered by implantation (for example subcutaneously orintramuscularly) or by intramuscular injection. Thus, for example, thecompounds may be formulated with suitable polymeric or hydrophobicmaterials (for example as an emulsion in an acceptable oil) or ionexchange resins, or as sparingly soluble derivatives, for example, as asparingly soluble salt.

For buccal or sublingual administration, the compositions may take theform of tablets, lozenges, pastilles, or gels formulated in conventionalmanner. Such compositions may comprise the active ingredient in aflavored basis such as sucrose and acacia or tragacanth.

The compounds may also be formulated in rectal compositions such assuppositories or retention enemas, e.g., containing conventionalsuppository bases such as cocoa butter, polyethylene glycol, or otherglycerides.

Certain compounds disclosed herein may be administered topically, thatis by non-systemic administration. This includes the application of acompound disclosed herein externally to the epidermis or the buccalcavity and the instillation of such a compound into the ear, eye andnose, such that the compound does not significantly enter the bloodstream. In contrast, systemic administration refers to oral,intravenous, intraperitoneal and intramuscular administration.

Formulations suitable for topical administration include liquid orsemi-liquid preparations suitable for penetration through the skin tothe site of inflammation such as gels, liniments, lotions, creams,ointments or pastes, and drops suitable for administration to the eye,ear or nose. The active ingredient for topical administration maycomprise, for example, from 0.001% to 10% w/w (by weight) of theformulation. In certain embodiments, the active ingredient may compriseas much as 10% w/w. In other embodiments, it may comprise less than 5%w/w. In certain embodiments, the active ingredient may comprise from 2%w/w to 5% w/w. In other embodiments, it may comprise from 0.1% to 1% w/wof the formulation.

For administration by inhalation, compounds may be convenientlydelivered from an insufflator, nebulizer pressurized packs or otherconvenient means of delivering an aerosol spray. Pressurized packs maycomprise a suitable propellant such as dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In the case of a pressurized aerosol, the dosageunit may be determined by providing a valve to deliver a metered amount.Alternatively, for administration by inhalation or insufflation, thecompounds may take the form of a dry powder composition, for example apowder mix of the compound and a suitable powder base such as lactose orstarch. The powder composition may be presented in unit dosage form, infor example, capsules, cartridges, gelatin or blister packs from whichthe powder may be administered with the aid of an inhalator orinsufflator.

In one embodiment, a compound is prepared for delivery in asustained-release, controlled release, extended-release, timed-releaseor delayed-release formulation, for example, in semipermeable matricesof solid hydrophobic polymers containing the therapeutic agent. Varioustypes of sustained-release materials have been established and are wellknown by those skilled in the art. Current extended-release formulationsinclude film-coated tablets, multiparticulate or pellet systems, matrixtechnologies using hydrophilic or lipophilic materials and wax-basedtablets with pore-forming excipients (see, for example, Huang, et al.Drug Dev. Ind. Pharm. 29:79 (2003); Pearnchob, et al. Drug Dev. Ind.Pharm. 29:925 (2003); Maggi, et al. Eur. J. Pharm. Biopharm. 55:99(2003); Khanvilkar, et al., Drug Dev. Ind. Pharm. 228:601 (2002); andSchmidt, et al., Int. J. Pharm. 216:9 (2001)). Sustained-releasedelivery systems can, depending on their design, release the compoundsover the course of hours or days, for instance, over 4, 6, 8, 10, 12,16, 20, 24 hours or more. Usually, sustained release formulations can beprepared using naturally-occurring or synthetic polymers, for instance,polymeric vinyl pyrrolidones, such as polyvinyl pyrrolidone (PVP);carboxyvinyl hydrophilic polymers; hydrophobic and/or hydrophilichydrocolloids, such as methylcellulose, ethylcellulose,hydroxypropylcellulose, and hydroxypropylmethylcellulose; andcarboxypolymethylene.

The sustained or extended-release formulations can also be preparedusing natural ingredients, such as minerals, including titanium dioxide,silicon dioxide, zinc oxide, and clay (see, U.S. Pat. No. 6,638,521,herein incorporated by reference). Exemplified extended releaseformulations that can be used in delivering a compound include thosedescribed in U.S. Pat. Nos. 6,635,680; 6,624,200; 6,613,361; 6,613,358,6,596,308; 6,589,563; 6,562,375; 6,548,084; 6,541,020; 6,537,579;6,528,080 and 6,524,621, each of which is hereby incorporated herein byreference. Controlled release formulations of particular interestinclude those described in U.S. Pat. Nos. 6,607,751; 6,599,529;6,569,463; 6,565,883; 6,482,440; 6,403,597; 6,319,919; 6,150,354;6,080,736; 5,672,356; 5,472,704; 5,445,829; 5,312,817 and 5,296,483,each of which is hereby incorporated herein by reference. Those skilledin the art will readily recognize other applicable sustained releaseformulations.

Systemic administration can also be by transmucosal or transdermalmeans. For transmucosal or transdermal administration, penetrantsappropriate to the barrier to be permeated are used in the formulation.For topical administration, the agents can be formulated into ointments,creams, salves, powders or gels. In one embodiment, the transdermaldelivery agent can be DMSO. Transdermal delivery systems can include,e.g., patches. For transmucosal administration, penetrants appropriateto the barrier to be permeated are used in the formulation. Suchpenetrants are generally known in the art. Exemplified transdermaldelivery formulations that can find use with the compounds disclosedherein include those described in U.S. Pat. Nos. 6,589,549; 6,544,548;6,517,864; 6,512,010; 6,465,006; 6,379,696; 6,312,717 and 6,310,177,each of which are hereby incorporated herein by reference.

The precise amount of compound administered to a patient will be theresponsibility of the attendant physician. The specific dose level forany particular patient will depend upon a variety of factors includingthe activity of the specific compound employed, the age, body weight,general health, sex, diets, time of administration, route ofadministration, rate of excretion, drug combination, the precisedisorder being treated, and the severity of the indication or conditionbeing treated. Also, the route of administration may vary depending onthe condition and its severity. The dosage can be increased or decreasedover time, as required by an individual patient. A patient initially maybe given a low dose, which is then increased to an efficacious dosagetolerable to the patient. Typically, a useful dosage for adults may befrom 5 to 2000 mg, but have been known to range from 0.1 to 500 mg/kgper day. By way of example, a dose may range from 1 to 200 mg, whenadministered by oral route; or from 0.1 to 100 mg or, in certainembodiments, 1 to 30 mg, when administered by intravenous route; in eachcase administered, for example, from 1 to 4 times per day. When acompound is administered in combination with another therapeutic agent,a useful dosage of the combination partner may be from 20% to 100% ofthe normally recommended dose, since, as discussed below, even doses ofa given drug which would be subtherapeutic if administered on its ownmay be therapeutic when used in combination with another agent.

Dosage amount and interval can be adjusted individually to provideplasma levels of the active compounds which are sufficient to maintaintherapeutic effect. In certain embodiments, therapeutically effectiveserum levels will be achieved by administering single daily doses, butefficacious multiple daily dose schedules may be used as well. In casesof local administration or selective uptake, the effective localconcentration of the drug may not be related to plasma concentration.One having skill in the art will be able to optimize therapeuticallyeffective local dosages without undue experimentation. Additionally,applicable methods for determining an appropriate dose and dosingschedule for administration of compounds such as those disclosed hereinare described, for example, in Goodman and Gilman's The PharmacologicalBasis of Therapeutics, 11^(th) Ed., Brunton, Lazo and Parker, Eds.,McGraw-Hill (2006), and in Remington: The Science and Practice ofPharmacy, 2E′ Ed., Gennaro, Ed., Lippencott Williams & Wilkins (2003),both of which are hereby incorporated herein by reference.

In certain instances, it may be appropriate to administer at least oneof the compounds described herein (or a pharmaceutically acceptablesalt, ester, or prodrug thereof) in combination with another therapeuticagent. By way of example only, if one of the side effects experienced bya patient upon receiving one of the compounds herein is hypertension,then it may be appropriate to administer an anti-hypertensive agent incombination with the initial therapeutic agent. Or, by way of exampleonly, the therapeutic effectiveness of one of the compounds describedherein may be enhanced by administration of an adjuvant (i.e., by itselfthe adjuvant may only have minimal therapeutic benefit, but incombination with another therapeutic agent, the overall therapeuticbenefit to the patient is enhanced). Or, by way of example only, thebenefit of experienced by a patient may be increased by administeringone of the compounds described herein with another therapeutic agent(which also includes a therapeutic regimen) that also has therapeuticbenefit. By way of example only, in a treatment for HIV dementiainvolving administration of one of the compounds described herein,increased therapeutic benefit may result by also providing the patientwith another therapeutic agent for dementia or inflammation. In anycase, regardless of the disease, disorder or condition being treated,the overall benefit experienced by the patient may simply be additive ofthe two therapeutic agents or the patient may experience a synergisticbenefit.

Specific, non-limiting examples of possible combination therapiesinclude use of certain compounds disclosed herein with compounds usedfor treating diseases and conditions which can be affected by SGLTinhibition, such as antidiabetic agents, lipid-lowering/lipid-modulatingagents, agents for treating diabetic complications, anti-obesity agents,antihypertensive agents, antihyperuricemic agents, and agents fortreating chronic heart failure, atherosclerosis or related disorders.

In any case, the multiple therapeutic agents (at least one of which is acompound disclosed herein) may be administered in any order or evensimultaneously. If simultaneously, the multiple therapeutic agents maybe provided in a single, unified form, or in multiple forms (by way ofexample only, either as a single pill or as two separate pills). One ofthe therapeutic agents may be given in multiple doses, or both may begiven as multiple doses. If not simultaneous, the timing between themultiple doses may be any duration of time ranging from a few minutes tofour weeks.

Examples of agents to be used in combination with compounds disclosedherein include lithium, valproate and other agents used inneuroprotection, PAF receptor antagonists, antioxidants includingmitochondrially-targeted antioxidants, activators of SIRT1 and othersirtuins, inhibitors of indoleamine 2,3 dehydrogenase (IDO), agentswhich enhance trans-blood brain bather (BBB) uptake of drugs, includingcompounds that inhibit drug pumps at the BBB such as, for example,ritonavir; HAART drugs and other agents for use in HIV treatment; agentsfor the treatment of cardiovascular, heart, and metabolic disorders,such as HMG-CoA reductase inhibitors including statins, insulin andinsulin mimetics, and glycogen synthase kinase-3 beta (GSK3β)inhibitors; agents which “normalize” mitochondrial function;antiinflammatory agents including PAF receptor antagonists or PAFacetylhydrolase, cyclooxygenase inhibitors (including COX-2 selectiveand nonselective) such as aspirin, ibuprofen, naproxen, and celecoxib;and agents for blocking liver cell proliferation, such as JNKinhibitors.

Also provided are combinations of multiple agents, such as lithium plusa GSK3β blocker, to be used in combination with the compounds providedherein.

Additionally, agents for neuroprotection and/or neurogenesis includeselective serotonin reuptake inhibitors SSRIs and small moleculeagonists of neurotrophin receptors.

Any of the aforementioned agents may be combined with viral vectors thatexpress genes intended to induce neural progenitor cells, as well.

Treatment with the compounds disclosed here in may also be effectivewhen delivered along with deep-brain stimulation, such as inParkinsonism and HIV-associated dementia/HIV-associated neurocognitivedisorder.

Thus, in another aspect, certain embodiments provide methods fortreating MLK3-mediated disorders in a human or animal subject in need ofsuch treatment comprising administering to said subject an amount of acompound disclosed herein effective to reduce or prevent said disorderin the subject, in combination with at least one additional agent forthe treatment of said disorder that is known in the art. In a relatedaspect, certain embodiments provide therapeutic compositions comprisingat least one compound disclosed herein in combination with one or moreadditional agents for the treatment of MLK3-mediated disorders.

Specific diseases to be treated by the compounds, compositions, andmethods disclosed herein include: metabolic diseases such as type 1 andtype 2 diabetes mellitus, hyperglycemia, diabetic complications (such asretinopathy, nephropathy, neuropathy, ulcers, micro- andmacroangiopathies, gout and diabetic foot disease), insulin resistance,metabolic syndrome (Syndrome X), hyperinsulinemia, hypertension,hyperuricemia, obesity, edema, dyslipidemia, hepatic steatosis,non-alcoholic steatohepatitis (NASH), chronic heart failure, andatherosclerosis.

Compounds disclosed herein may also be useful for the treatment ofinflammatory diseases such as bacterial sepsis, otitis media,endotoxemia, mucosal hyperplasia, inflammatory bowel disease, Crohn'sdisease, irritable bowel syndrome, and ulcerative colitis; andrespiratory diseases and conditions such as asthma, chronic obstructivepulmonary disease (COPD), and acute inhalation-induced lung injury.

Compounds disclosed herein may also be useful for the treatment ofautoimmune diseases such as multiple sclerosis, rheumatoid arthritis,lupus and Crohn's disease.

Compounds disclosed herein may also be useful for the treatment ofproliferative disorders including cancers such as liver cancer.Furthermore, Compounds disclosed herein may also be useful for thetreatment of hepatitis, including viral hepatitis, and non-alcoholicsteatohepatitis (NASH).

Compounds disclosed herein may also be useful for the treatment ofischemic injury, including stroke, cerebral ischemia/reperfusion,myocardial infarction, and ischemic heart disease.

Compounds disclosed herein may also be useful for the treatment ofdiseases and disorders of the nervous system such as Alzheimer's Disease(AD), Parkinson's Disease, HIV dementia, HIV associated neurocognitivedisorder (HAND), neuroinflammatory diseases, and neuropathies includingdrug-induced peripheral neuropathy, and diabetic neuropathy, andHIV-associated neuropathy, ototoxicity and hearing loss, acute insultsto the inner ear, including acoustic trauma, blast noise (for example,as experienced by military personnel), exposure to ototoxicchemotherapeutic agents for cancer therapy (such as cisplatin) andtreatment with aminoglycoside antibiotics. Compounds disclosed hereinmay also be useful for the treatment of traumatic brain injury includingstroke.

Compounds disclosed herein may also be useful for the treatment of painincluding inflammatory pain, neuropathic pain, back pain includingdiscogenic pain, the pain of arthritis and autoimmune disorders such asrheumatoid arthritis, and cancer pain including pain due to bonemetastasis.

Compounds disclosed herein may also be useful for the treatment ofpsychological disorders including depression or major depressivedisorder (MDD), bipolar disorder, and post-traumatic stress disorder.

Compounds disclosed herein may also be useful for enhancement ofstem-cell based therapies in the central nervous system (CNS).

All publications and patent applications cited in this specification areherein incorporated by reference as if each individual publication orpatent application were specifically and individually indicated to beincorporated by reference. Although the foregoing has been described insome detail by way of illustration and example for purposes of clarityof understanding, it will be readily apparent to those of ordinary skillin the art in light of the teachings of this invention that certainchanges and modifications can be made thereto without departing from thespirit or scope of the invention.

EXAMPLES

The invention is further illustrated by the following examples, whichare offered for illustrative purposes, and are not intended to limit theinvention in any manner. Those of skill in the art will readilyrecognize a variety of noncritical parameters, which can be changed ormodified to yield essentially the same results.

The structures of compounds synthesized in the examples below wereconfirmed using the following procedures. LC-MS/UV/ELS analysis wasperformed on instrumentation consisting of Shimadzu LC-LOAD vp seriesHPLC pumps and dual wavelength UV detector, a Gilson 215 autosampler, aSedex 75c evaporative light scattering (ELS) detector, and a PE/SciexAPI 150EX mass spectrometer. The ELS detector was set to a temperatureof 40° C., a gain setting of 7, and a N₂ pressure of 3.3 atm. The TurboIonSpray source was employed on the API 150 with an ion spray voltage of5 kV, a temperature of 300° C., and orifice and ring voltages of 5 V and175 V respectively. Positive ions were scanned in Q1 from 160 to 650m/z. 5.0 μL injections were performed for each sample, on a PhenomenexGemini 5 μm C18 column. Mobile phases consisted of 0.05% formic acid inboth HPLC grade water (A) and HPLC grade acetonitrile (B). 5.0 μLinjections were performed for each sample, using gradient elution from5% B to 100% B in 4 min at a flow rate of 2.0 mL/min with a final holdat 100% B of 1.8 min UV and ELS data is collected for 4.5 min Routineone-dimensional NMR spectroscopy was performed on a 300 MHz VarianMercury-Plus spectrometer. The samples were dissolved in deuteratedsolvents obtained from Cambridge Isotope Laboratories, Inc., andtransferred to 5 mm ID NMR tubes. The spectra were acquired at 293 K.The chemical shifts were recorded on the ppm scale and were referencedto the appropriate solvent signals, such as 2.49 ppm for DMSO-d6, 1.93ppm for CD₃CN, 3.30 ppm for CD₃OD, 5.32 ppm for CD₂Cl₂ and 7.26 ppm forCDCl₃ for ¹H spectra.

Other equipment and techniques standard in the art of chemical analysisand characterization may be used.

Example 1

Preparation of 5-bromo-3-iodo-1-tosyl-1H-pyrrolo[2,3-b]pyridine(Intermediate A)

To a stirred solution of 5-bromo-3-iodo-1H-pyrrolo[2,3-b]pyridine (0.70g, 2.2 mmol) in 15 mL of anhydrous THF cooled to 0° C. with an ice bathwas added NaH [60% dispersion in mineral oil] (0.13 g, 3.3 mmol). Thereaction mixture was stirred for 20 min at 0° C., after whichp-toluenesulfonyl chloride (0.47 g, 2.4 mmol) was added. The resultingmixture was stirred at 0° C. for 1.5 hr, after which cold 0.5 M HCl (20mL) was added. The mixture was partitioned between EtOAc and 0.5 M HCl,after which the organic layer was separated, dried over MgSO₄, filtered,and evaporated in vacuo to yield a residue that was triturated with 20%CH₂Cl₂ in hexanes to yield the title compound (0.84 g, 81%) as a lightyellow powder. ¹H NMR (DMSO-d6, 300 MHz) δ 8.51 (d, J=2.1 Hz, 1H), 8.22(s, 1H), 8.02 (d, J=1.2 Hz, 1H), 8.00 (d, J=5.1 Hz, 2H), 7.44 (dd, J=8.7Hz, 0.6 Hz, 2H), 2.35 (s, 3H); MS ESI (m/z): 477.0/479.0 (M+1)⁺, calc.476.

Preparation of5-bromo-3-(1H-indol-5-yl)-1-tosyl-1H-pyrrolo[2,3-b]pyridine(Intermediate B)

To a stirred suspension of5-bromo-3-iodo-1-tosyl-1H-pyrrolo[2,3-b]pyridine (0.35 g, 0.73 mmol) and1H-indol-5-ylboronic acid (0.14 mg, 0.88 mmol) in CH₃CN (10 mL) wasadded 1 M Na₂CO₃ (10 mL) followed bybis(triphenylphosphine)palladium(II)dichloride (0.050 g, 0.071 mmol).The resulting mixture was stirred overnight at 60° C. After the mixturewas evaporated to dryness in vacuo, it was dissolved in DMF (3 mL),absorbed onto Celite, and dried. The residue was purified via silica gelchromatography using CH₂Cl₂ as the eluent to obtain the title compound(0.26 g, 76%). ¹H NMR (CDCl₃, 300 MHz): δ 8.48 (d, J=2.1 Hz, 1H), 8.27(bs, 1H), 8.26 (d, J=2.4 Hz, 1H), 8.08 (d, J=8.1 Hz), 7.85 (s, 1H), 7.81(m, 1H), 7.50 (d, J=8.7 Hz, 1H), 7.37 (dd, J=1.8, 8.4 Hz), 7.30 (m, 3H),6.63 (m, 1H), 2.39 (s, 3H); MS ESI (m/z): 466.2/468.2 (M+1)⁺, calc. 465.

Preparation of3-(1H-indol-5-yl)-5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridine(Compound C)

To a solution of5-bromo-3-(1H-indol-5-yl)-1-tosyl-1H-pyrrolo[2,3-b]pyridine (65 mg, 0.14mmol) in CH₃CN (1 mL) in a Personal Chemistry microwave reaction vialwas added 3,4,5-trimethoxyphenylboronic acid (30 mg, 0.14 mmol),bis(triphenylphosphine)-palladium(II)dichloride (7.0 mg, 0.010 mmol),and 1 M Na₂CO₃ (1 mL). The resulting mixture was de-gassed with Ar for10 min, after which it was heated at 150° C. for 10 min in a PersonalChemistry Optimizer. The organic layer was separated, filtered, andconcentrated in vacuo. The residue was dissolved in MeOH (3 mL) andacetone (2 mL), and 2 M NaOH (1.5 mL) was added. The resulting mixturewas stirred at 65° C. for 30 min, after which it was partitioned betweenEtOAc and 1 M NaOH. The organic layer was separated, dried over MgSO₄,filtered, and stripped to give a residue purified via preparatory HPLCto give the title compound as a white solid. ¹H NMR (DMSO-d6, 300 MHz):δ 11.78 (s, 1H), 11.03 (s, 1H), 8.51 (d, J=2.1 Hz, 1H), 8.36 (d, J=1.8Hz, 1H), 7.86 (s, 1H), 7.72 (d, J=2.4 Hz, 1H), 7.45 (s, 2H), 7.32 (m,1H), 6.92 (s, 2H), 6.45 (m, 1H), 3.85 (s, 6H), 3.70 (s, 3H); HPLCretention time: 2.04 minutes; MS ESI (m/z): 400.4 (M+1)⁺, calc. 399.

Example 2 Preparation of5-(3,4-dimethoxyphenyl)-3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridine(Compound D)

Compound D was prepared by a method analogous to that described inExample 1 by substituting 3,4-dimethoxyphenylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate B.HPLC retention time: 2.33 minutes. MS ESI (m/z): 370.2 (M+H)⁺, calc.369.

Example 3 Preparation ofN-(4-(3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl)phenyl)acetamide(Compound E)

Compound E was prepared by a method analogous to that described inExample 1 by substituting 4-acetamidophenylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate B.HPLC retention time: 1.86 minutes. MS ESI (m/z): 367.4 (M+H)⁺, calc.366.

Example 4 Preparation of5-(3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl)pyridin-2-amine(Compound F)

Compound F was prepared by a method analogous to that described inExample 1 by substituting5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate B.¹H NMR (DMSO-d6, 300 MHz): δ 11.73 (d, J=1.8 Hz, 1H), 11.05 (s, 1H),8.43 (d, J=2.4 Hz, 1H), 8.29 (d, J=1.8 Hz, 1H), 8.27 (d, J=2.1 Hz, 1H),7.88 (s, 1H), 7.76 (dd, J=2.4, 8.4 Hz, 1H), 7.46 (s, 2H), 7.33 (m, 1H),6.55 (dd, J=0.6, 8.7 Hz, 1H), 6.46 (m, 1H), 5.99 (s, 2H). HPLC retentiontime: 1.10 minutes. MS ESI (m/z): 326.2 (M+H)⁺, calc. 325.

Example 5 Preparation of4-(3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl)-2-methoxyaniline(Compound G)

Compound G was prepared by a method analogous to that described inExample 1 by substituting 4-amino-3-methoxyphenylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate B.HPLC retention time: 1.54 minutes. MS ESI (m/z): 355.4 (M+H)⁺, calc.354.

Example 6 Preparation of3-(1H-indol-5-yl)-5-(6-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridine(Compound H)

Compound H was prepared by a method analogous to that described inExample 1 by substituting 6-methoxypyridin-3-ylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate B.HPLC retention time: 2.16 minutes. MS ESI (m/z): 341.4 (M+H)⁺, calc.340.

Example 7 Preparation of3-(1H-indol-5-yl)-5-(2-(4-methylpiperazin-1-yl)pyridin-4-yl)-1H-pyrrolo[2,3-b]pyridine(Compound I)

Compound I was prepared by a method analogous to that described inExample 1 by substituting 2-(4-methylpiperazin-1-yl)pyridin-4-ylboronicacid for 3,4,5-trimethoxyphenylboronic acid in the reaction withintermediate B. HPLC retention time: 1.37 minutes. MS ESI (m/z): 409.4(M+H)⁺, calc. 408.

Example 8 Preparation of4-(3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl)aniline (Compound J)

Compound J was prepared by a method analogous to that described inExample 1 by substituting 4-aminophenylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate B.HPLC retention time: 1.47 minutes. MS ESI (m/z): 325.4 (M+H)⁺, calc.324.

Example 9 Preparation of5-(3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl)pyrimidin-2-amine(Compound K)

Compound K was prepared by a method analogous to that described inExample 1 by substituting 2-aminopyrimidin-5-ylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate B.HPLC retention time: 1.81 minutes. MS ESI (m/z): 327.2 (M+H)⁺, calc.326.

Example 10 Preparation of3-(1H-indol-5-yl)-5-(6-(piperazin-1-yl)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridine(Compound L)

Compound L was prepared by a method analogous to that described inExample 1 by substituting 6-(piperazin-1-yl)pyridin-3-ylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate B.HPLC retention time: 1.15 minutes. MS ESI (m/z) 395.4 (M+H)⁺, calc. 394.

Example 11 Preparation ofN-(4-(3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl)phenyl)methanesulfonamide(Compound M)

Compound M was prepared by a method analogous to that described inExample 1 by substituting 4-(methylsulfonamido)phenylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate B.HPLC retention time: 1.99 minutes. MS ESI (m/z): 403.4 (M+H)⁺, calc.402.

Example 12 Preparation of3,5-di(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridine (Compound N)

Compound N was prepared by a method analogous to that described inExample 1 by substituting 1H-indol-5-ylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate B.HPLC retention time: 2.01 minutes. MS ESI (m/z): 349.2 (M+H)⁺, calc.348.

Example 13 Preparation of5-(3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl)-N,N-dimethylpyridin-2-amine(Compound O)

Compound O was prepared by a method analogous to that described inExample 1 by substituting 6-(dimethylamino)pyridin-3-ylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate B.HPLC retention time: 1.58 minutes. MS ESI (m/z): 354.4 (M+H)⁺, calc.353.

Example 14 Preparation of3-(1H-indol-5-yl)-5-phenyl-1H-pyrrolo[2,3-b]pyridine (Compound P)

Compound P was prepared by a method analogous to that described inExample 1 by substituting phenylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate B.HPLC retention time: 2.49 minutes. MS ESI (m/z): 310.2 (M+H)⁺, calc.309.

Example 15 Preparation of4-(5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)aniline(Compound Q)

Compound Q was prepared by a method analogous to that described inExample 1 by substituting 4-aminophenylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A. HPLCretention time: 1.45 minutes. MS ESI (m/z): 376.4 (M+H)⁺, calc. 375.

Example 16 Preparation ofN-(4-(5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)phenyl)acetamide(Compound R)

Compound R was prepared by a method analogous to that described inExample 1 by substituting 4-acetamidophenylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A. HPLCretention time: 1.98 minutes. MS ESI (m/z): 418.6 (M+H)⁺, calc. 417.

Example 17 Preparation of5-(5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)pyrimidin-2-amine(Compound S)

Compound S was prepared by a method analogous to that described inExample 1 by substituting 2-aminopyrimidin-5-ylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A. HPLCretention time: 1.98 minutes. MS ESI (m/z): 378.4 (M+H)⁺, calc. 377.

Example 18 Preparation of5-(5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-amine(Compound T)

Compound T was prepared by a method analogous to that described inExample 1 by substituting5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine for1H-indol-5-ylboronic acid in the reaction with Intermediate A. ¹H NMR(DMSO-d6, 300 MHz): δ 11.82 (s, 1H), 8.53 (d, J=1.8 Hz, 1H), 8.31 (d,J=1.8, 1H), 8.28 (d, J=1.5 Hz), 7.76 (dd, J=2.1, 8.4 Hz, 1H), 7.70 (d,J=2.4 Hz, 1H), 6.95 (s, 2H), 6.54 (d, J=8.4 Hz, 1H), 5.87 (s, 2H), 3.86(s, 6H), 3.68 (s, 3H); HPLC retention time: 1.10 minutes. MS ESI (m/z):377.4 (M+H)⁺, calc. 376.

Example 19 Preparation ofN,N-dimethyl-5-(5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-amine(Compound U)

Compound U was prepared by a method analogous to that described inExample 1 by substituting 6-(dimethylamino)pyridin-3-ylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A. HPLCretention time: 1.43 minutes. MS ESI (m/z): 405.6 (M+H)⁺, calc. 404.

Example 20 Preparation of5,5′-(1H-pyrrolo[2,3-b]pyridine-3,5-diyl)dipyrimidin-2-amine (CompoundW)

Compound W was prepared by a method analogous to that described inExample 1 by substituting 2-aminopyrimidin-5-ylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A and2-aminopyrimidin-5-ylboronic acid for 3,4,5-trimethoxyphenylboronic acidin the reaction with Intermediate B. HPLC retention time: 1.17 minutes.MS ESI (m/z): 305.2 (M+H)⁺, calc. 304.

Example 21 Preparation of5,5′-(1H-pyrrolo[2,3-b]pyridine-3,5-diyl)bis(N,N-dimethylpyridin-2-amine)(Compound X)

Compound X was prepared by a method analogous to that described inExample 1 by substituting 6-(dimethylamino)pyridin-3-ylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A and6-(dimethylamino)pyridin-3-ylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with Intermediate B.HPLC retention time: 1.17 minutes. MS ESI (m/z): 359.4 (M+H)⁺, calc.358.

Example 22 Preparation of5-(3-(3-chloro-4-fluorophenyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)-N,N-dimethylpyridin-2-amine(Compound Y)

Compound Y was prepared by a method analogous to that described inExample 1 by substituting 3-chloro-4-fluorophenylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A and6-(dimethylamino)pyridin-3-ylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with Intermediate B.HPLC retention time: 1.73 minutes. MS ESI (m/z): 367.2 (M+H)⁺, calc.366.

Example 23 Preparation of5-(3-(4-aminophenyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)pyridin-2-amine(Compound Z)

Compound Z was prepared by a method analogous to that described inExample 1 by substituting 4-aminophenylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A and6-aminopyridin-3-ylboronic acid for 3,4,5-trimethoxyphenylboronic acidin the reaction with Intermediate B. HPLC retention time: 0.68 minutes.MS ESI (m/z): 302.4 (M+H)⁺, calc. 301.

Example 24 Preparation of3-(1-methyl-1H-indol-5-yl)-5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridine(Compound AA)

Compound AA was prepared by a method analogous to that described inExample 1 by substituting 1-methyl-1H-indol-5-ylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A. HPLCretention time: 2.29 minutes. MS ESI (m/z): 414.4 (M+H)⁺, calc. 413.

Example 25 Preparation of4-(5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)benzamide(Compound AB)

Compound AB was prepared by a method analogous to that described inExample 1 by substituting 4-carbamoylphenylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A. HPLCretention time: 1.64 minutes. MS ESI (m/z): 404.6 (M+H)⁺, calc. 403.

Example 26 Preparation of4-(5-(3,4-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)benzamide(Compound AC)

Compound AC was prepared by a method analogous to that described inExample 1 by substituting 4-carbamoylphenylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A and3,4-dimethoxyphenylboronic acid for 3,4,5-trimethoxyphenylboronic acidin the reaction with Intermediate B. HPLC retention time: 1.60 minutes.MS ESI (m/z): 374.2 (M+H)⁺, calc. 373.

Example 27 Preparation of4-(5-(4-amino-3-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)benzamide(Compound AD)

Compound AD was prepared by a method analogous to that described inExample 1 by substituting 4-carbamoylphenylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A and4-amino-3-methoxyphenylboronic acid for 3,4,5-trimethoxyphenylboronicacid in the reaction with Intermediate B. HPLC retention time: 1.46minutes. MS ESI (m/z): 359.2 (M+H)⁺, calc. 358.

Example 28 Preparation of4-(5-(6-aminopyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)benzamide(Compound AE)

Compound AE was prepared by a method analogous to that described inExample 1 by substituting 4-carbamoylphenylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine for3,4,5-trimethoxyphenylboronic acid in the reaction with Intermediate B.HPLC retention time: 1.13 minutes. MS ESI (m/z): 330.4 (M+H)⁺, calc.329.

Example 29 Preparation of5-(3-(3-chloro-4-fluorophenyl)-1H-pyrrolo[2,3-b]pyridin-5-yl)pyridin-2-amine(Compound AF)

Compound AF was prepared by a method analogous to that described inExample 1 by substituting 3-chloro-4-fluorophenylboronic acid for1H-indol-5-ylboronic acid in the reaction with Intermediate A and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine for3,4,5-trimethoxyphenylboronic acid in the reaction with Intermediate B.HPLC retention time: 1.47 minutes. MS ESI (m/z): 339.4 (M+H)⁺, calc.338.

Example 30

Preparation of5-(3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl)-N-methylpyridin-2-amine(Compound All)

To a solution of5-bromo-3-(1H-indol-5-yl)-1-tosyl-1H-pyrrolo[2,3-b]pyridine (40 mg, 0.09mmol) in CH₃CN (1 mL) in a Personal Chemistry microwave reaction vialwas added 6-fluoropyridin-3-ylboronic acid (12 mg, 0.09 mmol),bis(triphenylphosphine)-palladium(II)dichloride (5.0 mg, 0.007 mmol),and 1 M Na₂CO₃ (1 mL). The resulting mixture was de-gassed with Ar for10 min, after which it was heated at 150° C. for 10 min in a PersonalChemistry Optimizer. The organic layer was separated, filtered, andconcentrated in vacuo to give intermediate Q. The residue was dissolvedin DMSO (0.5 mL) and methylamine hydrochloride salt (29 mg, 0.43 mmol),and K₂CO₃ (95 mg, 0.70 mmol) were added. The resulting mixture wasstirred at 80° C. for 48 hr, after which it was diluted with DMF (0.5mL), filtered, and subjected to preparative HPLC to yield the titlecompound (6.0 mg, 21%). ¹H NMR (DMSO-d6, 300 MHz): δ 11.77 (s, 1H),11.07 (s, 1H), 8.46 (d, J=2.1 Hz, 1H), 8.34 (dd, J=2.4, 9.3 Hz, 1H),7.90 (s, 1H), 7.86 (m, 1H), 7.74 (d, J=2.7 Hz, 1H), 7.47 (s, 2H), 7.35(s, 1H), 6.80 (s, 1H), 6.63 (d, J=8.4 Hz, 1H), 6.48 (m, 1H), 2.84 (d,J=4.5 Hz, 1H). HPLC retention time: 1.10 minutes; HPLC retention time:1.56 minutes; MS ESI (m/z): 340.2 (M+1)⁺, calc. 339.

Example 31 Preparation of5-(3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl)-N-(2-(pyrrolidin-1-yl)ethyl)pyridin-2-amine(Compound AI)

Compound AI was prepared by a method analogous to that described inExample 15 by substituting 2-(pyrrolidin-1-yl)ethanamine for methylaminehydrochloride salt in the reaction with intermediate Q. HPLC retentiontime: 1.58 minutes. MS ESI (m/z): 354.4 (M+H)⁺, calc. 353.

Example 32

Preparation of4-(3-(1H-indol-5-yl)-1-tosyl-1H-pyrrolo[2,3-b]pyridin-5-yl)benzaldehyde(Intermediate AJ)

To a solution of5-bromo-3-(1H-indol-5-yl)-1-tosyl-1H-pyrrolo[2,3-b]pyridine[Intermediate B] (0.20 g, 0.43 mmole) in CH₃CN (4 mL) in a PersonalChemistry microwave reaction vial was added 4-formylphenylboronic acid(64 mg, 0.43 mmol), bis(triphenylphosphine)-palladium(II)dichloride (40mg, 0.057 mmol), and 1 M Na₂CO₃ (2 mL). The resulting mixture wasde-gassed with Ar for 10 min, after which it was heated at 150° C. for10 min in a Personal Chemistry Optimizer. The organic layer wasseparated, filtered, and partitioned between EtOAc and brine. Theorganic layer was dried over MgSO₄, filtered, and concentrated in vacuoto give Intermediate AJ. HPLC retention time: 3.01 minutes. MS ESI(m/z): 492.4 (M+H)⁺, calc. 491.

Preparation of3-(1H-indol-5-yl)-5-(4-((4-methylpiperazin-1-yl)methyl)phenyl)-1H-pyrrolo[2,3-b]pyridine(Compound AK)

To a solution of4-(3-(1H-indol-5-yl)-1-tosyl-1H-pyrrolo[2,3-b]pyridin-5-yl)benzaldehyde[Intermediate AJ] (0.11 g, 0.214 mmol) in CH₂Cl₂ (3 mL) was added1-methylpiperazine (40 μL, 0.40 mmol) and sodium triacetoxyborohydride(68 mg, 0.32 mmol). The reaction mixture was stirred for 1 hr at roomtemperature, after which it was partitioned between CH₂Cl₂ and 1 M NaOH.The organic layer was separated, dried over MgSO₄, and concentrated invacuo. The residue was dissolved in 3:2 MeOH:acetone (5 mL), and 2 MNaOH (1.5 mL) was added. The resulting mixture was stirred at 65° C. for30 min, after which it was partitioned between EtOAc and 1 M NaOH. Theorganic layer was separated, dried over MgSO₄, filtered, and stripped toprovide a residue that was subjected to preparatory HPLC to yield thetitle compound. HPLC retention time: 1.63 minutes; MS ESI (m/z) 422.4(M+1)⁺, calc. 421.

Example 33 Preparation of1-(4-(3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl)phenyl)-N,N-dimethylmethanamine(Compound AL)

Compound AL was prepared by a method analogous to that described inExample 33 by substituting dimethylamine (2 M solution in THF) for1-methylpiperazine in the reaction with intermediate T. HPLC retentiontime: 1.66 minutes. MS ESI (m/z): 367.4 (M+H)⁺, calc. 366.

Example 34

Preparation of1-(2-(2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethyl)piperazine(Intermediate AM)

To a solution of 2-(piperazin-1-yl)ethanol (0.78 mL, 6.0 mmol) andtriphenylphosphine (1.6 g, 6.0 mmol) in anhydrous THF (20 mL) at 0° C.was added diethyl azodicarboxylate (0.95 mL, 6.0 mmol), followed by2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-phenol (1.0 g,4.0 mmol). After stiffing for 4 h at rt, additional triphenylphosphine(1.6 g, 6.0 mmol) and diethyl azodicarboxylate (0.95 mL, 6.0 mmol) wereadded. After stirring for an additional 2 h, the resulting mixture wasevaporated to dryness in vacuo and the residue was purified via silicagel chromatography eluting with 15% MeOH in CH₂Cl₂ to yield a yellow oil(1.89 g) which contained approximately 60% of the title compound by HPLCanalysis. HPLC retention time: 1.01 minutes. MS ESI (m/z): 363.6 (M+H)⁺,calc. 362.

Preparation of3-(1H-indol-5-yl)-5-(3-methoxy-4-(2-(piperazin-1-yl)ethoxy)phenyl)-1H-pyrrolo[2,3-b]pyridine(Compound AO)

To a solution of5-bromo-3-(1H-indol-5-yl)-1-tosyl-1H-pyrrolo[2,3-b]pyridine(Intermediate B) (92 mg, 0.20 mmol) in CH₃CN (2 mL) in a PersonalChemistry microwave reaction vial was added1-(2-(2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethyl)piperazine(Intermediate AM) (72 mg, 0.20 mmol),bis(triphenylphosphine)-palladium(II)dichloride (20 mg, 0.028 mmol), and1 M Na₂CO₃ (2 mL). The resulting mixture was de-gassed with Ar for 10min, after which it was heated at 150° C. for 25 min in a PersonalChemistry Optimizer. The organic layer was separated, filtered, andconcentrated in vacuo to give Intermediate AN. The residue was dissolvedin MeOH (3 mL) and acetone (2 mL), and 2 M NaOH (1.5 mL) was added. Theresulting mixture was stirred at 50° C. for 2 h, after which it waspartitioned between EtOAc and 1 M NaOH. The organic layer was separated,dried over MgSO₄, filtered, and stripped to give a residue that wassubjected to preparatory HPLC to yield the title compound. HPLCretention time: 1.29 minutes; MS ESI (m/z) 468.6 (M+1)⁺, calc. 467.

Example 35 Preparation of2-(4-(3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl)-2-methoxyphenoxy)-N,N-dimethylethanamine(Compound AP)

Compound AP was prepared by a method analogous to that described inExample 36 by substituting 2-(dimethylamino)ethanol for2-(piperazin-1-yl)ethanol in the reaction with2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol. HPLCretention time: 1.20 minutes. MS ESI (m/z): 427.2 (M+H)⁺, calc. 426.

Example 36

Preparation of 5-bromo-3-((trimethylsilyl)ethynyl)pyrazin-2-amine(Intermediate AQ)

To a solution of 3,5-dibromopyrazin-2-amine (10 g, 40 mmol), copper(I)iodide (0.91 g, 4.7 mmol), diisopropylethylamine (53 mL, 0.55 mol), andtetrakis(triphenylphosphine)-palladium(0) (2.3 g, 1.9 mmol) in DMF (120mL) that was de-gassed with Ar was added trimethylsilylacetylene (6.7mL, 48 mmol). The resulting mixture was stirred under an Ar atmospherefor 1 h at 120° C., after which it was evaporated to dryness in vacuo.The residue was subjected to silica gel chromatography eluting with 35%EtOAc in hexanes to give a brown oil that was triturated with hexanes togive the title compound (5.0 g, 47%). ¹H NMR (CDCl₃, 300 MHz): δ 8.04(s, 1H), 5.10 (s, 2H), 0.28 (s, 9H). HPLC retention time: 2.75 minutes.MS ESI (m/z): 270.0, 272.0 (M+H)⁺, calc. 269.

Preparation ofN-(5-bromo-3-((trimethylsilyl)ethynyl)pyrazin-2-yl)acetamide(Intermediate AR)

To a solution of 5-bromo-3-((trimethylsilyl)ethynyl)pyrazin-2-amine (5.0g, 19 mmol) and pyridine (3.8 mL, 46 mmol) in anhydrous THF (75 mL) wasadded acetyl chloride (1.6 mL, 23 mmol) in a drop-wise manner. Afterstirring for 48 hr at rt, additional acetyl chloride (0.4 mL, 6 mmol)was added and the mixture was stirred for an additional 48 hr at rt. Thesolvent was removed in vacuo, and the residue was diluted with 30% EtOAcin hexanes. The mixture was filtered, and the filtrate was purified viasilica gel chromatography eluting with 30% EtOAc in hexanes to give ayellow-brown solid (1.8 g, 31%). ¹H NMR (CDCl₃, 300 MHz): δ 8.34 (s,1H), 8.08 (s, 1H), 2.46 (s, 3H), 0.32 (s, 9H). HPLC retention time: 2.29minutes. MS ESI (m/z): 312.2, 314.2 (M+H)⁺, calc. 311.

Preparation of 2-bromo-5H-pyrrolo[3,2-b]pyrazine (Intermediate AS)

A solution ofN-(5-bromo-3-((trimethylsilyl)ethynyl)pyrazin-2-yl)acetamide[Intermediate AR] (2.6 g, 8.4 mmol) and tetrabutylammonium fluoride [1 Min THF] (18 mL, 18 mmol) in anhydrous THF (26 mL) was heated at 75° C.for 20 h, after which it was partitioned between EtOAc and H₂O. Theorganic layer was washed with brine, dried over Na₂SO₄, and evaporatedin vacuo to yield a residue that was purified via silica gelchromatography eluting with 30% EtOAc in hexanes to give the titlecompound as a tan solid (0.69 g, 42%). ¹H NMR (CDCl₃, 300 MHz): δ 8.88(bs, 1H), 8.34 (s, 1H), 7.62 (t, J=3.3 Hz, 1H), 6.71 (dd, J=3.6 Hz, 3.9Hz, 1H). HPLC retention time: 1.73 minutes. MS ESI (m/z): 198.2, 200.2(M+H)⁺, calc. 197.

Preparation of 2-bromo-7-iodo-5H-pyrrolo[3,2-b]pyrazine (IntermediateAT)

To a solution of 2-bromo-5H-pyrrolo[3,2-b]pyrazine [Intermediate AS](0.68 g, 3.4 mmol) in acetone (17 mL) was added N-iodosuccinimide (0.82g, 3.6 mmol) and the resulting mixture was stirred for 4 h at rt. Themixture was evaporated in vacuo to yield a residue that was purified viasilica gel chromatography eluting with 40% THF in hexanes to give thetitle compound as a yellow solid (0.99 g, 89%). ¹H NMR (DMSO-d6, 300MHz): δ 12.82 (s, 1H), 8.42 (s, 1H), 8.20 (s, 1H). HPLC retention time:2.23 minutes. MS ESI (m/z): 324.0, 326.0 (M+H)⁺, calc. 323.

Preparation of 2-bromo-7-iodo-5-tosyl-5H-pyrrolo[3,2-b]pyrazine(Intermediate AU)

To a stirred solution of 2-bromo-7-iodo-5H-pyrrolo[3,2-b]pyrazine[Intermediate AT] (1.1 g, 3.5 mmol) in anhydrous THF (20 mL) cooled to0° C. was added NaH [60% dispersion in mineral oil] (0.17 g, 4.3 mmol).The reaction mixture was stirred for 20 min at 0° C., after whichp-toluenesulfonyl chloride (0.73 g, 3.8 mmol) in THF (8 mL) was added.The resulting mixture was stirred at rt for 3 hr, after which it wasdiluted with EtOAc and washed with H₂O and brine. The organic layer wasseparated, dried over Na₂SO₄, filtered, and evaporated in vacuo to yielda residue that was triturated with hexanes to yield the title compound(1.6 g, 94%) as a light yellow powder. ¹H NMR (DMSO-d6, 300 MHz) δ 8.62(d, J=7.5 Hz, 2H), 8.03 (s, 1H), 8.00 (s, 1H), 7.47 (d, J=8.1 Hz, 2H),2.37 (s, 3H). HPLC retention time: 2.84 minutes. MS ESI (m/z):478.0/480.0 (M+H)⁺, calc. 477.

Preparation of2-bromo-7-(1H-indol-5-yl)-5-tosyl-5H-pyrrolo[3,2-b]pyrazine(Intermediate AV)

To a stirred suspension of2-bromo-7-iodo-5-tosyl-5H-pyrrolo[3,2-b]pyrazine [Intermediate AU] (0.25g, 0.52 mmol) and 1H-indol-5-ylboronic acid (0.10 mg, 0.62 mmol) inCH₃CN (20 mL) was added 1 M Na₂CO₃ (20 mL) followed bybis(triphenylphosphine)-palladium(II)dichloride (60 mg, 0.086 mmol). Theresulting mixture was stirred for 2 h at 60° C. The title compound wasisolated as a yellow solid via filtration from the CH₃CN layer (0.23 g,94%). HPLC retention time: 3.23 minutes. MS ESI (m/z): 467.2/469.2(M+H)⁺, calc. 466.

Preparation of7-(1H-indol-5-yl)-2-(3,4,5-trimethoxyphenyl)-5H-pyrrolo[3,2-b]pyrazine(Compound AW)

To a solution of2-bromo-7-(1H-indol-5-yl)-5-tosl-5H-pyrrolo[3,2-b]pyrazine [IntermediateAV] (65 mg, 0.14 mmol) in CH₃CN (1 mL) in a Personal Chemistry microwavereaction vial was added 3,4,5-trimethoxyphenylboronic acid (30 mg, 0.14mmol), bis(triphenylphosphine)-palladium(II)dichloride (7.0 mg, 0.010mmol), and 1 M Na₂CO₃ (1 mL). The resulting mixture was de-gassed withAr for 10 min, after which it was heated at 150° C. for 10 min in aPersonal Chemistry Optimizer. The organic layer was separated, filtered,and concentrated in vacuo. The residue was dissolved in MeOH (3 mL) andacetone (2 mL), and 2 M NaOH (1.5 mL) was added. The resulting mixturewas stirred at 65° C. for 30 min, after which it was partitioned betweenEtOAc and 1 M NaOH. The organic layer was separated, dried over MgSO₄,filtered, and stripped to give a residue which was purified bypreparatory HPLC to give the title compound as a yellow solid. HPLCretention time: 2.25 minutes; MS ESI (m/z) 401.2 (M+1)⁺, calc. 400.

Example 37 Preparation of2-(3,4-dimethoxyphenyl)-7-(1H-indol-5-yl)-5H-pyrrolo[3,2-b]pyrazine(Compound AX)

Compound AX was prepared by a method analogous to that described inExample 38 by substituting 3,4-dimethoxyboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate AV.HPLC retention time: 2.45 minutes. MS ESI (m/z): 371.2 (M+H)⁺, calc.370.

Example 38 Preparation of4-(7-(1H-indol-5-yl)-5H-pyrrolo[3,2-b]pyrazin-2-yl)-2-methoxyaniline(Compound AY)

Compound AY was prepared by a method analogous to that described inExample 38 by substituting 4-amino-3-methoxyphenylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with intermediate AV.HPLC retention time: 2.07 minutes. MS ESI (m/z): 356.4 (M+H)⁺, calc.355.

Example 39 Preparation of4-(2-(4-(7-(1H-indol-5-yl)-5H-pyrrolo[3,2-b]pyrazin-2-yl)-2-methoxyphenoxy)ethyl)morpholine(Compound AZ)

Compound AZ was prepared by a method analogous to that described inExample 36 by substituting4-(2-(2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethyl)morpholinefor1-(2-(2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethyl)piperazineand 2-bromo-7-(1H-indol-5-yl)-5-tosyl-5H-pyrrolo[3,2-b]pyrazine forintermediate B. HPLC retention time: 1.59 minutes. MS ESI (m/z): 470.4(M+H)⁺, calc. 469.

Example 40

Preparation of 3,5-dibromopyrazin-2-amine (Intermediate BA)

To a stirred solution of aminopyrazine (8.21 g, 86.4 mmol) in anhydrousmethylene chloride (215 mL) cooled to 0° C. was added N-bromosuccinimide(32.3 g, 181 mmol) in portions over a six hour period, during which timethe temperature of the reaction was kept below 0° C. The resultingmixture was stored at 4° C. overnight, after which it was stirredvigorously and quenched with H₂O (100 mL). The organic layer wasseparated, after which it was washed with saturated aqueous NaHCO₃,washed with brine, dried over MgSO₄, filtered, and evaporated in vacuoto yield a residue that was triturated with 20% EtOAc in hexanes toyield the title compound (10.3 g, 47%) as a yellow/brown powder. ¹H NMR(CDCl₃, 300 MHz) δ 8.02 (s, 1H), 5.05 (bs, 2H); HPLC retention time:1.99 minutes; MS ESI (m/z): 252.0/254.0/256.2 (M+1)⁺, calc. 251.

Preparation of 6-bromo-N²-(1H-indol-5-yl)pyrazine-2,3-diamine(Intermediate BB)

To a stirred suspension of 3,5-dibromopyrazin-2-amine (3.48 g, 13.7mmol) and 1H-indol-5-amine (2.00 g, 15.0 mmol) in EtOH (3.5 mL) wasadded diisopropylethylamine [DIEA] (2.60 mL, 15.0 mmol). The resultingmixture was stirred for 48 hr at 80° C., after which it was partitionedbetween EtOAc and H₂O. The organic layer was separated, after which itwas washed with brine, dried over Na₂SO₄, filtered, and evaporated invacuo to yield a residue that was purified via silica gel chromatographyeluting with 1:1 EtOAc:hexanes to yield the title compound (1.75 g, 42%)as a red/brown solid. ¹H NMR (DMSO-d6, 300 MHz): δ 10.98 (s, 1H), 8.22(s, 1H), 7.83 (s, 1H), 7.31-7.28 (m, 3H), 7.19 (d, J=8.7 Hz, 1H), 6.43(s, 2H), 6.36 (s, 1H); HPLC retention time: 2.07 minutes; MS ESI (m/z):304.2/306.2 (M+1)⁺, calc. 303.

Preparation of6-bromo-1-(1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one(Intermediate BC)

To a solution of 6-bromo-N²-(1H-indol-5-yl)pyrazine-2,3-diamine (0.450g, 1.48 mmol) in THF (5 mL) was added carbonyldiimidazole (1.20 g, 7.40mmol). The resulting mixture was heated at 65° C. for 48 hr, after whichit was concentrated in vacuo and partitioned between EtOAc and H₂O. Theorganic layer was separated, dried over MgSO₄, filtered, andconcentrated in vacuo to yield a residue that was purified via silicagel chromatography eluting with EtOAc to yield the title compound (0.20g, 41%). HPLC retention time: 2.07 minutes; MS ESI (m/z): 330.2/332.2(M+1)⁺, calc. 329.

Preparation of1-(1H-indol-5-yl)-6-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one(Compound BD)

To a solution of6-bromo-1-(1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one (27 mg,0.08 mmol) in CH₃CN (1 mL) in a Personal Chemistry microwave reactionvial was added 3,4,5-trimethoxyphenylboronic acid (17 mg, 0.08 mmol),bis(triphenylphosphine)-palladium(II)dichloride (6.0 mg, 0.008 mmol),and 1 M Na₂CO₃ (1 mL). The resulting mixture was de-gassed with Ar for10 min, after which it was heated at 150° C. for 10 min in a PersonalChemistry Optimizer. The organic layer was separated, filtered, andconcentrated in vacuo. The residue was purified by preparatory HPLC toyield the title compound (6.5 mg, 19%). ¹H NMR (DMSO-d6, 300 MHz): δ12.18 (s, 1H), 11.28 (s, 1H), 8.57 (s, 1H), 7.83 (d, J=1.8 Hz, 1H), 7.52(d, J=8.4 Hz, 1H), 7.42 (m, 1H), 7.37 (dd, J=1.8, 8.4 Hz, 1H), 7.20 (s,2H), 6.51 (m, 1H), 3.78 (s, 6H), 3.66 (s, 3H); HPLC retention time: 2.30minutes; MS ESI (m/z): 418.4 (M+1)⁺, calc. 417.

Example 41 Preparation of1-(1-methyl-1H-indol-5-yl)-6-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one(Compound BE)

Compound BE was prepared by a method analogous to that described inExample 1 by substituting 1-methyl-1H-indol-5-amine for 1H-indol-5-aminein the reaction with Intermediate BA. 4.0 mg recovered. ¹H NMR (DMSO-d6,300 MHz): δ 12.22 (s, 1H), 8.57 (s, 1H), 7.85 (d, J=1.8 Hz, 1H), 7.57(d, J=8.7 Hz, 1H), 7.45 (d, J=1.8 Hz), 7.41 (m, 2H), 7.20 (s, 2H), 6.50(d, J=3.0 Hz, 1H), 3.84 (s, 3H), 3.78 (s, 6H), 3.66 (s, 3H); HPLCretention time: 2.50 minutes. MS ESI (m/z): 432.4 (M+H)⁺, calc. 431.

Example 42 Preparation of6-(4-hydroxyphenyl)-1-(1-methyl-1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one(Compound BF)

Compound BE was prepared by a method analogous to that described inExample 1 by substituting 1-methyl-1H-indol-5-amine for 1H-indol-5-aminein the reaction with Intermediate BA to prepare6-bromo-1-(1-methyl-1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one.In a procedure similar to that used to synthesize Compound D,4-hydroxyphenylboronic acid was substituted for3,4,5-trimethoxyphenylboronic acid and6-bromo-1-(1-methyl-1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-onewas substituted for6-bromo-1-(1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one to obtainthe title compound. 2.2 mg recovered. HPLC retention time: 2.18 minutes.MS ESI (m/z): 358.2 (M+H)⁺, calc. 357.

Example 43 Preparation of6-(3,5-dimethylphenyl)-1-(1-methyl-1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one(Compound BG)

Compound BG was prepared by a method analogous to that described inExample 3 by substituting 3,5-dimethylphenylboronic acid for4-hydroxyphenylboronic acid in the reaction with6-bromo-1-(1-methyl-1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one.1.6 mg recovered. HPLC retention time: 3.04 minutes. MS ESI (m/z): 370.2(M+H)⁺, calc. 369.

Example 44 Preparation of1-(1H-indol-5-yl)-6-(pyridin-4-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one(Compound BH)

Compound BH was prepared by a method analogous to that described inExample 1 by substituting pyridin-4-ylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with Compound BC. 1.6mg recovered. HPLC retention time: 1.10 minutes. MS ESI (m/z): 329.4(M+H)⁺, calc. 328.

Example 45 Preparation of6-(4-hydroxyphenyl)-1-(1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one(Compound BI)

Compound BI was prepared by a method analogous to that described inExample 1 by substituting by substituting 4-hydroxyphenylboronic acidfor 3,4,5-trimethoxyphenylboronic acid in the reaction with Compound BC.13.7 mg recovered. ¹H NMR (DMSO-d6, 300 MHz): δ 12.07 (s, 1H), 11.30 (s,1H), 9.61 (s, 1H), 8.38 (s, 1H), 7.69 (m, 2H), 7.52 (d, J=8.4 Hz, 1H),7.44 (m, 1H), 7.26 (dd, J=1.8, 8.7 Hz), 6.76 (dd, J=2.4, 12.9 Hz), 6.52(m, 1H); HPLC retention time: 1.99 minutes. MS ESI (m/z): 344.2 (M+H)⁺,calc. 343.

Example 46 Preparation of6-(3,5-dimethylphenyl)-1-(1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one(Compound BJ)

Compound BJ was prepared by a method analogous to that described inExample 1 by substituting 3,5-dimethylphenylboronic acid for3,4,5-trimethoxyphenylboronic acid in the reaction with Compound BC. 4.3mg recovered. HPLC retention time: 2.80 minutes. MS ESI (m/z): 356.2(M+H)⁺, calc. 355.

Examples 47-119

Examples 47-119, shown in Table 3 below, were synthesized in parallelaccording to procedures given below in Schemes 7 and 8, using thereagents in Tables 1 and 2.

Examples 47-67

Preparation of 1H-imidazo[4,5-b]pyrazin-2(3H)-one compounds in Table 1

To a solution of6-bromo-1-(1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one (27 mg,0.08 mmol) in CH₃CN (1 mL) in a Personal Chemistry microwave reactionvial was added 3,4,5-trimethoxyphenylboronic acid (17 mg, 0.08 mmol),bis(triphenylphosphine)-palladium(II)dichloride (6.0 mg, 0.008 mmol),and 1 M Na₂CO₃ (1 mL). The resulting mixture was de-gassed with Ar for10 min, after which it was heated at 150° C. for 10 min in a PersonalChemistry Optimizer. The organic layer was separated, filtered, andconcentrated in vacuo. The residue was purified by preparatory HPLC toyield the title compounds (>3 mg) in table 1, isolated as amorphoussolids.

TABLE 1 Ex. Boronic Acid Purified Compound Isolated 473,4-dimethoxyphenyl 6-(3,4-dimethoxyphenyl)-1-(1H-indol-5- boronic acidyl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 48 3,5-dichlorophenyl boronic6-(3,5-dichlorophenyl)-1-(1H-indol-5- acidyl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 49 3-fluoro-4-methoxyphenyl6-(3-fluoro-4-methoxyphenyl)-1-(1H- boronic acidindol-5-yl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 503-amino-4-methoxyphenyl 6-(3-amino-4-methoxyphenyl)-1-(1H- boronic acidindol-5-yl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 51 4-methoxy-3,5-1-(1H-indol-5-yl)-6-(4-methoxy-3,5- dimethylphenyl boronicdimethylphenyl)-1H-imidazo[4,5- acid b]pyrazin-2(3H)-one 524-morpholinophenyl 1-(1H-indol-5-yl)-6-(4- boronic acidmorpholinophenyl)-1H-imidazo[4,5- b]pyrazin-2(3H)-one 53Indole-5-boronic acid 1,6-di(1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 54 3-hydroxyphenyl boronic6-(3-hydroxyphenyl)-1-(1H-indol-5-yl)- acid1H-imidazo[4,5-b]pyrazin-2(3H)-one 55 4-hydroxy-3-6-(4-hydroxy-3-methoxyphenyl)-1-(1H- methoxyphenylindol-5-yl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 56 indole-6-boronic1-(1H-indol-5-yl)-6-(1H-indol-6-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one57 3-methoxy-4-(2- 1-(1H-indol-5-yl)-6-(3-methoxy-4-(2-morpholinoethoxy)phenyl morpholinoethoxy)phenyl)-1H- boronic acidimidazo[4,5-b]pyrazin-2(3H)-one 58 2,5-difluoro-4-6-(2,5-difluoro-4-hydroxyphenyl)-1-(1H- hydroxyphenyl boronicindol-5-yl)-1H-imidazo[4,5-b]pyrazin- acid 2(3H)-one 59 3,5-difluoro-4-6-(3,5-difluoro-4-hydroxyphenyl)-1-(1H- hydroxyphenyl boronicindol-5-yl)-1H-imidazo[4,5-b]pyrazin- acid 2(3H)-one 604-amino-3-methoxyphenyl 6-(4-amino-3-methoxyphenyl)-1-(1H- boronic acidindol-5-yl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 61 3,5-difluorophenyl6-(3,5-difluorophenyl)-1-(1H-indol-5- boronic acidyl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 62 4-hydroxy-3,5-6-(4-hydroxy-3,5-dimethoxyphenyl)-1- dimethoxyphenyl(1H-indol-5-yl)-1H-imidazo[4,5- boronic acid b]pyrazin-2(3H)-one 63 2,3-6-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)- dihydrobenzo[b][1,4]dioxin-1-(1H-indol-5-yl)-1H-imidazo[4,5- 6-boronic acid b]pyrazin-2(3H)-one 644-hydroxy-3,5- 6-(4-hydroxy-3,5-dimethylphenyl)-1- dimethylphenylboronic (1H-indol-5-yl)-1H-imidazo[4,5- acid b]pyrazin-2(3H)-one 653,5-dimethoxyphenyl 6-(3,5-dimethoxyphenyl)-1-(1H-indol-5- boronic acidyl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 66 2-(4-methylpiperazin-1-1-(1H-indol-5-yl)-6-(2-(4- yl)pyridin-4-boronic acidmethylpiperazin-1-yl)pyridin-4-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one67 (3-methoxy-4-(2- 1-(1H-indol-5-yl)-6-(3-methoxy-4-(2- (piperazin-1-(piperazin-1-yl)ethoxy)phenyl)-1H- yl)ethoxy)phenylimidazo[4,5-b]pyrazin-2(3H)-one

Examples 68-118

Preparation of Intermediates 2BB

To a stirred suspension of 3,5-dibromopyrazin-2-amine (3.48 g, 13.7mmol) and the corresponding alkyl, aryl, or heteroaryl amine (15.0 mmol)in EtOH (3.5 mL) was added diisopropylethylamine [DIEA] (2.60 mL, 15.0mmol). The resulting mixture was stirred for 48 hr at 80° C., afterwhich it was partitioned between EtOAc and H₂O. The organic layer wasseparated, after which it was washed with brine, dried over Na₂SO₄,filtered, and evaporated in vacuo to yield a residue that was purifiedby automated medium pressure silica gel chromatography eluting with 1:1EtOAc:hexanes to yield the intermediates as amorphous solids.

Preparation of intermediates 2BC

Intermediates 2BB (0.450 g, 1.5 mmol) were dissolved in THF (5 mL) andtreated with carbonyldiimidazole (1.20 g, 7.40 mmol). The resultingmixture was heated at 65° C. for 48 hr, after which it was concentratedin vacuo and partitioned between EtOAc and H₂O. The organic layer wasseparated, dried over MgSO₄, filtered, and concentrated in vacuo toyield a residue that was purified via automated silica gelchromatography eluting with hexane/EtOAc to yield the intermediates 2B Cas amorphous solids.

Preparation of 1H-imidazo[4,5-b]pyrazin-2(3H)-one compounds in Table 2

Individual solutions of intermediates 2BC (0.08 mmol) in CH₃CN (1 mL) ina Personal Chemistry microwave reaction vial was added the correspondinglboronic acid (0.08 mmol),bis(triphenylphosphine)-palladium(II)dichloride (6.0 mg, 0.008 mmol),and 1 M Na₂CO₃ (1 mL). The resulting mixture was de-gassed with Ar for10 min, after which it was heated at 150° C. for 10 min in a PersonalChemistry Optimizer. The organic layer was separated, filtered, andconcentrated in vacuo. The residue was purified by preparatory HPLC toyield the title compounds in table 2 (>3 mg) as amorphous solids.

70 3,4- 4-methoxy- 6-(3,4-dimethoxyphenyl)-1- dimethoxyphenyl aniline(4-methoxyphenyl)-1H- boronic acid imidazo[4,5-b]pyrazin- 2(3H)-one 71pyridin-4-boronic 4-methoxy- 1-(4-methoxyphenyl)-6- acid boronic acidaniline (pyridin-4-yl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 72 3,4,5-2-methyl-5- 1-(2-methyl-1H-indol-5-yl)- trimethoxyphenyl amino-indole6-(3,4,5-trimethoxyphenyl)- boronic acid 1H-imidazo[4,5-b]pyrazin-2(3H)-one 73 3,5-dichlorophenyl 2-methyl-5- 6-(3,5-dichlorophenyl)-1-(2-boronic acid amino-indole methyl-1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 74 3,4,5- 1-amino-1-cyclopentyl-6-(3,4,5- trimethoxyphenyl cyclopentanetrimethoxyphenyl)-1H- boronic acid imidazo[4,5-b]pyrazin- 2(3H)-one 753,4- 1-amino- 1-cyclopentyl-6-(3,4- dimethoxyphenyl cyclopentanedimethoxyphenyl)-1H- boronic acid imidazo[4,5-b]pyrazin- 2(3H)-one 764-hydroxyphenyl 1-amino- 1-cyclopentyl-6-(4- boronic acid cyclopentanehydroxyphenyl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 77 pyridin-4-boronic1-amino- 1-cyclopentyl-6-(pyridin-4- acid cyclopentaneyl)-1H-imidazo[4,5- b]pyrazin-2(3H)-one 78 3,4,5- Cyclopropane-1-(cyclopropylmethyl)-6- trimethoxyphenyl methylamine(3,4,5-trimethoxyphenyl)- boronic acid 1H-imidazo[4,5-b]pyrazin-2(3H)-one 79 3,4- Cyclopropane- 1-(cyclopropylmethyl)-6- dimethoxyphenylmethylamine (3,4-dimethoxyphenyl)-1H- boronic acidimidazo[4,5-b]pyrazin- 2(3H)-one 80 3,5-dichlorophenyl Cyclopropane-1-(cyclopropylmethyl)-6- boronic acid methylamine(3,5-dichlorophenyl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 814-hydroxyphenyl Cyclopropane- 1-(cyclopropylmethyl)-6-(4- boronic acidmethylamine hydroxyphenyl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 824-aminopyridine Cyclopropane- 1-(cyclopropylmethyl)-6- boronic acidmethylamine (pyridin-4-yl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 833,4,5- 1H-Indazol-5- 1-(1H-indazol-5-yl)-6- trimethoxyphenyl amine(3,4,5-trimethoxyphenyl)- boronic acid 1H-imidazo[4,5-b]pyrazin-2(3H)-one 84 4-hydroxypheny 2-methyl-5- 6-(4-hydroxyphenyl)-1-(2-boronic acid amino-indole methyl-1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 85 pyridin-4-boronic 2-methyl-5-1-(2-methyl-1H-indol-5-yl)- acid boronic acid amino-indole6-(pyridin-4-yl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 864-morpholinophenyl Cyclopropane- 1-(cyclopropylmethyl)-6-(4- boronicacid methylamine morpholinophenyl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one87 3,4- 1H-Indazol-5- 6-(3,4-dimethoxyphenyl)-1- dimethoxyphenyl amine(1H-indazol-5-yl)-1H- boronic acid imidazo[4,5-b]pyrazin- 2(3H)-one 884-aminopyridine 1H-Indazol-5- 1-(1H-indazol-5-yl)-6- boronic acid amine(pyridin-4-yl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 894-morpholinophenyl 1H-Indazol-5- 1-(1H-indazol-5-yl)-6-(4- boronic acidamine morpholinophenyl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 90 3,4,5-1H-Indazol-5- 1-(1H-indazol-6-yl)-6- trimethoxyphenyl amine(3,4,5-trimethoxyphenyl)- boronic acid 1H-imidazo[4,5-b]pyrazin-2(3H)-one 91 3,4- 1H-Indazol-5- 6-(3,4-dimethoxyphenyl)-1-dimethoxyphenyl amine (1H-indazol-6-yl)-1H- boronic acidimidazo[4,5-b]pyrazin- 2(3H)-one 92 4-hydroxyphenyl 1H-Indazol-5-6-(4-hydroxyphenyl)-1-(1H- boronic acid amine indazol-6-yl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 93 4-aminopyridine 1H-Indazol-5-1-(1H-indazol-6-yl)-6- boronic acid amine (pyridin-4-yl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 94 2,4,6- 1-amino-1-cyclopentyl-6-(2,4,6- trimethoxyphenyl cyclopentanetrimethoxyphenyl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 953,5-dimethylphenyl 1H-Indazol-5- 6-(3,5-dimethylphenyl)-1- boronic acidamine (1H-indazol-6-yl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 96 3,4,5-benzo[d]thiazol- 1-(benzo[d]thiazol-5-yl)-6- trimethoxyphenyl 5-amine(3,4,5-trimethoxyphenyl)- boronic acid 1H-imidazo[4,5-b]pyrazin-2(3H)-one 97 4-hydroxyphenyl benzo[d]thiazol-1-(benzo[d]thiazol-5-yl)-6- boronic acid 5-amine (4-hydroxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 98 4-aminopyridine benzo[d]thiazol-1-(benzo[d]thiazol-5-yl)-6- boronic acid 5-amine (pyridin-4-yl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 99 3,5-dimethylphenyl benzo[d]thiazol-1-(benzo[d]thiazol-5-yl)-6- boronic acid 5-amine(3,5-dimethylphenyl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 1004-morpholinophenyl benzo[d]thiazol- 1-(benzo[d]thiazol-5-yl)-6- boronicacid 5-amine (4-morpholinophenyl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one101 3,4,5- 2,3-dihydro-1H- 1-(2,3-dihydro-1H-inden-1- trimethoxyphenylinden-1-amine yl)-6-(3,4,5- boronic acid trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 102 3,4,5- 1H-benzo[d]imidazol-1-(1H-benzo[d]imidazol-5- trimethoxyphenyl 5-amine yl)-6-(3,4,5- boronicacid trimethoxyphenyl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 103 3,4-1H-benzo[d]imidazol- 1-(1H-benzo[d]imidazol-5- dimethoxyphenyl 5-amineyl)-6-(3,4- boronic acid dimethoxyphenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 104 4-morpholinophenyl 1H-benzo[d]imidazol-1-(1H-benzo[d]imidazol-5- boronic acid 5-amineyl)-6-(4-morpholinophenyl)- 1H-imidazo[4,5-b]pyrazin- 2(3H)-one 1053,4,5- aniline 1-phenyl-6-(3,4,5- trimethoxyphenyl trimethoxyphenyl)-1H-boronic acid imidazo[4,5-b]pyrazin- 2(3H)-one 106 3,4- aniline6-(3,4-dimethoxyphenyl)-1- dimethoxyphenyl phenyl-1H-imidazo[4,5-boronic acid b]pyrazin-2(3H)-one 107 3-methoxy-4-(2- Cyclopropane-1-(cyclopropylmethyl)-6-(3- morpholino- methylamine methoxy-4-(2-ethoxy)phenyl morpholinoethoxy)phenyl)- boronic acid1H-imidazo[4,5-b]pyrazin- 2(3H)-one 108 3-methoxy-4-(2- 1-amino-1-cyclopentyl-6-(3-methoxy- morpholino- cyclopentane 4-(2- ethoxy)phenylmorpholinoethoxy)phenyl) - 1H-imidazo[4,5-b]pyrazin- 2(3H)-one 1093,4,5- 6- 1-(6-morpholinopyridin-3- trimethoxyphenyl morpholino-yl)-6-(3,4,5- boronic acid pyridin-3-amine trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 110 3,4,5- 2,3-dihydro-1H-1-(2,3-dihydro-1H-inden-2- trimethoxyphenyl inden-2-amine yl)-6-(3,4,5-boronic acid trimethoxyphenyl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 1113,4- 1H-pyrrolo[2,3- 6-(3,4-dimethoxyphenyl)-1- dimethoxyphenylb]pyridin-5- (1H-pyrrolo[2,3-b]pyridin-5- boronic acid amineyl)-1H-imidazo[4,5- b]pyrazin-2(3H)-one 112 3,4,5- 1H-pyrrolo[2,3-1-(1H-pyrrolo[2,3-b]pyridin- trimethoxyphenyl b]pyridin-5-5-yl)-6-(3,4,5- boronic acid amine trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 113 3,4,5- 1H-indol-6-1-(1H-indol-6-yl)-6-(3,4,5- trimethoxyphenyl amine trimethoxyphenyl)-1H-boronic acid imidazo[4,5-b]pyrazin- 2(3H)-one 114 3,4,5- 4-aminophenol1-(4-hydroxyphenyl)-6- trimethoxyphenyl (3,4,5-trimethoxyphenyl)-boronic acid 1H-imidazo[4,5-b]pyrazin- 2(3H)-one 115 3,4- 4-aminophenol6-(3,4-dimethoxyphenyl)-1- dimethoxyphenyl (4-hydroxyphenyl)-1H- boronicacid imidazo[4,5-b]pyrazin- 2(3H)-one 116 4-morpholinophenyl4-aminophenol 1-(4-hydroxyphenyl)-6-(4- boronic acidmorpholinophenyl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 1176-aminopyridin-3- 1-amino- 6-(6-aminopyridin-3-yl)-1- boronic acidcyclopentane cyclopentyl-1H-imidazo[4,5- b]pyrazin-2(3H)-one 1184-amino-3- 1-amino- 6-(4-amino-3- methoxyphenyl cyclopentanemethoxyphenyl)-1- boronic acid cyclopentyl-1H-imidazo[4,5-b]pyrazin-2(3H)-one

Examples 47-118 were physically characterized by electrospray ionizationmass spectrometry. Structures and molecular masses are given below inTable 3.

TABLE 3 Ex. Structure IUPAC Name MW  47

6-(3,4-dimethoxyphenyl)-1- (1H-indol-5-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 387.13  48

6-(3,5-dichlorophenyl)-1- (1H-indol-5-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 395.03  49

6-(3-fluoro-4- methoxyphenyl)-1-(1H- indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 375.11  50

6-(3-amino-4- methoxyphenyl)-1-(1H- indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 372.13  51

1-(1H-indol-5-yl)-6-(4- methoxy-3,5- dimethylphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 371.40  52

1-(1H-indol-5-yl)-6-(4- morpholinophenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 412.45  53

1,6-di(1H-indol-5-yl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one 366.39  54

6-(3-hydroxyphenyl)-1-(1H- indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 343.35  55

6-(4-hydroxy-3- methoxyphenyl)-1-(1H- indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 373.37  56

1-(1H-indol-5-yl)-6-(1H- indol-6-yl)-1H-imidazo[4,5- b]pyrazin-2(3H)-one366.39  57

1-(1H-indol-5-yl)-6-(3- methoxy-4-(2- morpholinoethoxy)phenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 486.53  58

6-(2,5-difluoro-4- hydroxyphenyl)-1-(1H- indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 379.33  59

6-(3,5-difluoro-4- hydroxyphenyl)-1-(1H- indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 379.33  60

6-(4-amino-3- methoxyphenyl)-1-(1H- indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 372.39  61

6-(3,5-difluorophenyl)-1- (1H-indol-5-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 363.33  62

6-(4-hydroxy-3,5- dimethoxyphenyl)-1-(1H- indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 403.40  63

6-(2,3- dihydrobenzo[b][1,4]dioxin- 6-yl)-1-(1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 385.39  64

6-(4-hydroxy-3,5- dimethylphenyl)-1-(1H- indol-5-yl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one 385.43  65

6-(3,5-dimethoxyphenyl)-1- (1H-indol-5-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 387.40  66

1-(1H-indol-5-yl)-6-(2-(4- methylpiperazin-1- yl)pyridin-4-yl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 426.50  67

1-(1H-indol-5-yl)-6-(3- methoxy-4-(2-(piperazin-1- yl)ethoxy)phenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 485.55  68

1-(4-methoxyphenyl)-6- (3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 408.42  69

6-(3,4-dimethoxyphenyl)-1- (4-methoxyphenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 378.39  70

6-(4-hydroxyphenyl)-1-(4- methoxyphenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 334.34  71

1-(4-methoxyphenyl)-6- (pyridin-4-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 319.33  72

1-(2-methoxy-1H-indol-5-yl)- 6-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 431.45  73

6-(3,5-dichlorophenyl)-1-(2- methyl-1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 410.27  74

1-cyclopentyl-6-(3,4,5- trimethoxyphenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 370.41  75

1-cyclopentyl-6-(3,4- dimethoxyphenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 340.39  76

1-cyclopentyl-6-(4- hydroxyphenyl)-1H- imidazo[4,5-b]pyrazin- 2(3H)-one296.33  77

1-cyclopentyl-6-(pyridin-4- yl)-1H-imidazo[4,5- b]pyrazin-2(3H)-one281.32  78

1-(cyclopropylmethyl)-6- (3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 356.38  79

1-(cyclopropylmethyl)-6- (3,4-dimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 326.36  80

1-(cyclopropylmethyl)-6- (3,5-dichlorophenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 335.20  81

1-(cyclopropylmethyl)-6-(4- hydroxyphenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 282.30  82

1-(cyclopropylmethyl)-6- (pyridin-4-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 267.29  83

1-(1H-imidazol-5-yl)-6- (3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 418.42  84

6-(4-hydroxyphenyl)-1-(2- methyl-1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 357.37  85

1-(2-methyl-1H-indol-5-yl)- 6-(pyridin-4-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 342.36  86

1-(cyclopropylmethyl)-6-(4- morpholinophenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 351.41  87

6-(3,4-dimethoxyphenyl)-1- (1H-indazol-5-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 388.39  88

1-(1H-indazol-5-yl)-6- (pyridin-4-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 329.32  89

1-(1H-indazol-5-yl)-6-(4- morpholinophenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 413.44  90

1-(1H-indazol-6-yl)-6- (3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 418.42  91

6-(3,4-dimethoxyphenyl)-1- (1H-indazol-6-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 388.39  92

6-(4-hydroxyphenyl)-1-(1H- indazol-6-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 344.34  93

1-(1H-indazol-6-yl)-6- (pyridin-4-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 329.32  94

1-cyclopentyl-6-(2,4,6- trimethoxyphenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 370.41  95

6-(3,5-dimethylphenyl)-1- (1H-indazol-6-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 356.39  96

1-(benzo[d]thiazol-5-yl)-6- (3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 435.46  97

1-(benzo[d]thiazol-5-yl)-6- (4-hydroxyphenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 361.38  98

1-(benzo[d]thiazol-5-yl)-6- (pyridin-4-yl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 346.37  99

1-(benzo[d]thiazol-5-yl)-6- (3,5-dimethylphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 373.44 100

1-(benzo[d]thiazol-5-yl)-6- (4-morpholinophenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 430.49 101

1-(2,3-dihydro-1H-inden-1- yl)-6-(3,4,5- trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 418.46 102

1-(1H-benzo[d]imidazol-5- yl)-6-(3,4,5- trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 418.42 103

1-(1H-benzo[d]imidazol-5- yl)-6-(3,4- dimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 388.39 104

1-(1H-benzo[d]imidazol-5- yl)-6-(4- morpholinophenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 413.44 105

1-phenyl-6-(3,4,5- trimethoxyphenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 378.39 106

6-(3,4-dimethoxyphenyl)-1- phenyl-1H-imidazo[4,5- b]pyrazin-2(3H)-one348.36 107

1-(cyclopropylmethyl)-6-(3- methoxy-4-(2- morpholinoethoxy)phenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 425.49 108

1-cyclopentyl-6-(3- methoxy-4-(2- morpholinoethoxy)phenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 439.52 109

1-(6-morpholinopyridin-3- yl)-6-(3,4,5- trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 464.48 110

1-(2,3-dihydro-1H-inden-2- yl)-6-(3,4,5- trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 418.46 111

6-(3,4-dimethoxyphenyl)-1- (1H-pyrrolo[2,3-b]pyridin-5-yl)-1H-imidazo[4,5- b]pyrazin-2(3H)-one 388.39 112

1-(1H-pyrrolo[2,3- b]pyridin-5-yl)-6-(3,4,5- trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 418.42 113

1-(1H-indol-6-yl)-6-(3,4,5- trimethoxyphenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 418.42 114

1-(4-hydroxyphenyl)-6- (3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin- 2(3H)-one 394.50 115

6-(3,4-dimethoxyphenyl)-1- (4-hydroxyphenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 364.50 116

1-(4-hydroxyphenyl)-6-(4- morpholinophenyl)-1H- imidazo[4,5-b]pyrazin-2(3H)-one 389.50 117

6-(6-aminopyridin-3-yl)-1- cyclopentyl-1H- imidazo[4,5-b]pyrazin-2(3H)-one 296.33 118

6-(4-amino-3- methoxyphenyl)-1- cyclopentyl-1H- imidazo[4,5-b]pyrazin-2(3H)-one 325.37

Example 119 Preparation of5-chloro-1-(cyclopropylmethyl)-6-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazin-2(3H)-one

Example 119 was prepared by a method analogous to that described inExamples 68-118 by substituting 6 chloro-3,5-dibromopyrazin-2-amine for3,5-dibromopyrazin-2-amine in the reaction with aminomethylcyclopropane.MS ESI (m/z): 390.83 calc

Example 120

Preparation of1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine(Intermediate BK)

To a stirred solution of 7-azaindole (1.18 g, 10.0 mmol) in anhydrousdimethylformamide (10 mL) cooled to 0° C. was added NaH [60% dispersionin mineral oil] (0.480 g, 12.0 mmol) in portions over 15 min. Theresulting mixture was allowed to stir for 1 hr at 0° C., after which(2-(chloromethoxy)ethyl)trimethylsilane [SEM-Cl] (2.12 mL, 12.0 mmol)was added over 15 min. The resulting mixture was stirred for 1 hr, afterwhich it was quenched with H₂O (50 mL), and partioned between EtOAc andH₂O. The organic layer was separated, washed with brine, dried overMgSO₄, filtered, and evaporated in vacuo to yield a yellow oil (2.50 g,100%). HPLC retention time: 2.66 minutes; MS ESI (m/z): 249.4 (M+1)⁺,calc. 248.

Preparation of3,3,5-tribromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-2(3H)-one(Compound BL)

To a solution of1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (2.50 g,10.0 mmol) in 1:1 tert-butanol/H₂O (140 mL) at room temperature wasadded bromine (6.40 mL, 126 mmol). After stiffing for 3.5 hr at roomtemperature, an additional portion of bromine was added (6.40 mL, 126mmol) and the resulting mixture was stirred for 18 hr. The resultingmixture was concentrated in vacuo to yield the title compound, which wasused without any further purification. HPLC retention time: 2.97minutes; MS ESI (m/z): 441.0/443.0/445.2 (Fragment+1)⁺, calc. 498.

Preparation of5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-2(3H)-one(Compound BM)

To a solution of3,3,5-tribromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-2(3H)-one(4.98 g, 10.0 mmol) in AcOH (50 mL) was added zinc dust (1.28 g, 20.0mmol). The resulting mixture was stirred at room temperature for 2 hr,after which it was filtered thru Celite and concentrated in vacuo. Theresulting residue was purified via silica gel chromatography elutingwith 1:1 Hexanes:EtOAc to yield the title compound as a yellow oil (0.85g, 25% over three steps). HPLC retention time: 2.60 minutes; MS ESI(m/z): 287.2 (Fragment+1)⁺, calc. 342.

Preparation of5-(3,4,5-trimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)-methyl)-1H-pyrrolo[2,3-b]pyridin-2(3H)-one(Compound BN)

To a solution of5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-2(3H)-one(0.85 g, 2.5 mmol) in CH₃CN (5 mL) was added3,4,5-trimethoxyphenylboronic acid (525 mg, 2.5 mmol),bis(triphenylphosphine)-palladium(II) dichloride (250 mg, 0.35 mmol),and 1 M Na₂CO₃ (5 mL). The resulting mixture was de-gassed with Ar for10 min, after which it was heated at 80° C. for 2 hr. The reactionmixture was partitioned between EtOAc and H₂O, and the organic layer wasseparated, filtered, and concentrated in vacuo. The residue was purifiedby silica gel chromatography eluting with 3:1 EtOAc:Hexanes to yield thetitle compound (640 mg, 60%). HPLC retention time: 2.51 minutes; MS ESI(m/z): 431.4 (M+1)⁺, calc. 430.

Preparation of3,3-dimethyl-5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2(3H)-one(Compound BO)

To a solution of5-(3,4,5-trimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)-methyl)-1H-pyrrolo[2,3-b]pyridin-2(3H)-one(43 mg, 0.10 mmol) in DMF (2 mL) was added cesium carbonate (0.17 g,0.50 mmol) and methyl iodide (19 μL 0.30 mmol). The resulting solutionwas stirred for 48 hr at room temperature, after which it waspartitioned between EtOAc and H₂O. The organic layer was separated,dried over MgSO₄, filtered, and concentrated in vacuo. The residue wasdissolved in 6 N HCl (10 mL) and MeOH (5 mL), and the resulting mixturewas stirred at room temperature overnight, after which it waspartitioned between EtOAc and H₂O. The organic layer was concentrated invacuo, and the residue was dissolved in EtOH (2 mL). Potassium acetate(100 mg) was then added, and the reaction was stirred for 2 hr. Theresulting solution was purified via preparatory HPLC to give the titlecompound (24 mg, 73%). ¹H NMR (CDCl₃, 300 MHz): δ 9.72 (s, 1H), 8.35 (d,J=2.1 Hz, 1H), 7.60 (d, J=1.8 Hz, 1H), 6.71 (s, 2H), 3.95 (s, 6H), 3.90(s, 3H), 1.49 (s, 6H). HPLC retention time: 1.80 minutes; MS ESI (m/z):329.4 (M+1)⁺, calc. 328.

Example 121

Preparation of5′-(3,4,5-trimethoxyphenyl)spiro[cyclopentane-1,3′-pyrrolo[2,3-b]pyridin]-2′(1′H)-one(Compound BP)

To a solution of5-(3,4,5-trimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)-methyl)-1H-pyrrolo[2,3-b]pyridin-2(3H)-one(Compound BN, 43 mg, 0.10 mmol) in DMF (2 mL) was added cesium carbonate(0.17 g, 0.50 mmol) and 1,4-diiodobutane (13 μL, 0.10 mmol). Theresulting solution was stirred for 4 hr at room temperature, after whichit was partitioned between EtOAc and H₂O. The organic layer wasseparated, dried over MgSO₄, filtered, and concentrated in vacuo. Theresidue was dissolved in 6 N HCl (10 mL) and MeOH (5 mL), and theresulting mixture was stirred at room temperature overnight, after whichit was partitioned between EtOAc and H₂O. The organic layer wasconcentrated in vacuo, and the residue was dissolved in EtOH (2 mL).Potassium acetate (100 mg) was then added, and the reaction was stirredfor 2 hr. The resulting solution was purified via preparatory HPLC togive the title compound (18 mg, 51%). ¹H NMR (CDCl₃, 300 MHz): δ 9.53(s, 1H), 8.32 (d, J=2.1 Hz, 1H), 7.56 (s, 1H), 6.69 (s, 2H), 3.95 (s,6H), 3.90 (s, 3H), 2.28 (m, 2H), 2.24 (m, 2H), 1.97 (m, 4H). HPLCretention time: 2.00 minutes; MS ESI (m/z): 355.4 (M+1)⁺, calc. 354.

Examples 122 and 123

Preparation of3,3-bis(cyclopropylmethyl)-5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2(3H)-one(Example 22, Compound BQ) and3-(cyclopropylmethyl)-5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2(3H)-one(Example 23, Compound BR)

To a solution of5-(3,4,5-trimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)-methyl)-1H-pyrrolo[2,3-b]pyridin-2(3H)-one(43 mg, 0.10 mmol) in DMF (2 mL) was added cesium carbonate (0.17 g,0.50 mmol), (bromomethyl)cyclopropane (10 μL, 0.10 mmol), and potassiumiodide (83 mg, 0.50 mmol). The resulting solution was stirred for 4 hrat room temperature, after which it was partitioned between EtOAc andH₂O. The organic layer was separated, dried over MgSO₄, filtered, andconcentrated in vacuo. The residue was dissolved in 6 N HCl (10 mL) andMeOH (5 mL), and the resulting mixture was stirred at room temperatureovernight, after which it was partitioned between EtOAc and H₂O. Theorganic layer was concentrated in vacuo, and the residue was dissolvedin EtOH (2 mL). Potassium acetate (100 mg) was then added, and thereaction was stirred for 2 hr. The resulting solution was purified viapreparatory HPLC to give the Compound Q (11.4 mg) and Compound R (4.1mg). Compound BQ: ¹H NMR (CDCl₃, 300 MHz): δ 8.37 (d, J=2.1 Hz, 1H),7.71 (s, 1H), 6.72 (s, 2H), 3.96 (s, 6H), 3.91 (s, 3H), 2.04 (m, 2H),1.69 (m, 2H), 1.26 (m, 2H), 0.88 (m, 2H), 0.40 (m, 2H), 0.29 (m, 2H),−0.07 (m, 2H). HPLC retention time: 2.49 minutes; MS ESI (m/z): 409.4(M+1)⁺, calc. 408. Compound BR: ¹H NMR (CDCl₃, 300 MHz): δ 8.31 (s, 1H),7.92 (s, 1H), 6.69 (s, 2H), 3.95 (s, 6H), 3.91 (s, 3H), 3.50 (m, 1H),2.18 (m, 1H), 1.78 (m, 1H), 1.26 (m, 1H), 0.83 (m, 2H), 0.25 (m, 2H).HPLC retention time: 2.32 minutes; MS ESI (m/z): 355.0 (M+1)⁺, calc.354.

Example 124

Preparation of1-(1H-indol-5-yl)-6-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyridin-2(3H)-one(Compound BS)

Commercially available 5-bromopyridine-2,3-diamine 3 was converted to6-bromo-1H-imidazo[4,5-b]pyridin-2(3H)-one via treatment with carbonyldiimidazole in THF at 60° C., which was then protected as the monoethoxycarbonyl derivative 4 in a fashion similar to that described in J. Org.Chem., 1995, 1565-1582. Intermediate 4 was subjected to an NOE analysis,and interactions between the 7-position hydrogen and the carbamate ethylgroup were apparent, supporting the structure that is shown above.Following protection of the 3-position amine with a tert-butylcarboxylate group and deprotection of the ethyl carboxylate group usingisopropyl amine, intermediate 6 was coupled to indole-5-boronic acidusing copper acetate in a mixture of DCM/pyridine, after which it wasdeprotected using TFA/CH₂Cl₂. To the resulting6-bromo-1-(1H-indol-5-yl)-1H-imidazo[4,5-b]pyridin-2(3H)-one in CH₃CN (1mL) in a microwave reaction vial was added 3,4,5-trimethoxyphenylboronicacid (30 mg, 0.14 mmol), bis(triphenylphosphine)-palladium(II)dichloride (7.0 mg, 0.010 mmol), and 1 M Na₂CO₃ (1 mL). The resultingmixture was de-gassed with Ar for 10 min, after which it was heated at150° C. for 10 min in a Personal Chemistry Optimizer. The resultingmixture was partitioned between EtOAc and 1 M NaOH. The organic layerwas separated, dried over MgSO₄, filtered, and stripped to give aresidue that was purified via preparatory HPLC to give 1.8 mg of thetitle compound. HPLC retention time: 2.36 minutes; MS ESI (m/z): 417.4(M+1)⁺, calc. 416.

Example 125 Preparation of1-(cyclopropylmethyl)-6-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyridin-2(3H)-one

Intermediate 5 from Example 124 was alkylated with(bromomethyl)cyclopropane using K₂CO₃ in acetone, after which it wasdeprotected using TFA/CH₂Cl₂. To the resulting6-bromo-1-(cyclopropylmethyl)-1H-imidazo[4,5-b]pyridin-2(3H)-one inCH₃CN (1 mL) in a microwave reaction vial was added3,4,5-trimethoxyphenylboronic acid (30 mg, 0.14 mmol),bis(triphenylphosphine)-palladium(II)dichloride (7.0 mg, 0.010 mmol),and 1 M Na₂CO₃ (1 mL). The resulting mixture was de-gassed with Ar for10 min, after which it was heated at 150° C. for 10 min in a PersonalChemistry Optimizer. The resulting mixture was partitioned between EtOAcand 1 M NaOH. The organic layer was separated, dried over MgSO₄,filtered, and stripped to give a residue that was purified viapreparatory HPLC to give 3.7 mg of the title compound. HPLC retentiontime: 1.90 minutes; MS ESI (m/z): 356.2 (M+1)⁺, calc. 355.

Example 126

Preparation of1-(1H-indol-5-yl)-6-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazine(Compound BW)

Following a method described in Pteridines, 2002, Vol. 13, 65-72,Intermediate BB was heated in anhydrous DMF at 175° C. for 15 min in aPersonal Chemistry Optimizer. To the resulting6-bromo-1-(1H-indol-5-yl)-1H-imidazo[4,5-b]pyrazine 1 in CH₃CN (1 mL) ina microwave reaction vial was added 3,4,5-trimethoxyphenylboronic acid(30 mg, 0.14 mmol), bis(triphenylphosphine)-palladium(II) dichloride(7.0 mg, 0.010 mmol), and 1 M Na₂CO₃ (1 mL). The resulting mixture wasde-gassed with Ar for 10 min, after which it was heated at 150° C. for10 min in a Personal Chemistry Optimizer. The resulting mixture waspartitioned between EtOAc and 1 M NaOH. The organic layer was separated,dried over MgSO₄, filtered, and stripped to give a residue that waspurified via preparatory HPLC to give 4.7 mg of the title compound. HPLCretention time: 2.43 minutes; MS ESI (m/z): 402.8 (M+1)⁺, calc. 401.

Example 127 Preparation of1-(cyclopropylmethyl)-6-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-b]pyrazine-2(3H)-thione(Compound BX)

Compound BX was prepared by reacting Example 78 with Lawesson's reagentin refluxing toluene. The resulting mixture was partitioned betweenEtOAc and 1 M NaHCO₃. The organic layer was separated, dried over MgSO₄,filtered, and stripped to give a residue that was purified viapreparatory HPLC to give 2.0 mg of the title compound. HPLC retentiontime: 2.29 minutes; MS ESI (m/z): 373.2 (M+1)⁺, calc. 372.

Example 128

Preparation of3-pyridin-3-ylmethylene-5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one

To a solution of5-(3,4,5-trimethoxy-phenyl)-1-(2-trimethylsilanyl-ethoxymethyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one(157 mg, 0.365 mmol) in toluene (2 mL) was added triethylamine (56 μl,0.365 mmol), molecular sieves 4 Å (100 mg), and 3-pyridinecarboxaldehyde(38 μl, 0.401 mmol). The resulting mixture was stirred overnight at roomtemperature, after which it was filtered and partitioned between DCM andH₂O. The organic layer was separated, dried over MgSO₄, filtered, andconcentrated in vacuo. The residue was purified by silica gelchromatography eluting with 40-70% EtOAc:Hexanes to yield theSEM-protected precursor as a mixture of cis and trans isomers (101 mg,53%). 41 mg (0.079 mmol) of this material was dissolved in MeOH (1.5ml), 6 N HCl (3 ml) was added, and the mixture was stirred for 3 hoursat 45° C. The reaction was quenched with 1 N NaOH (15 ml), neutralizedby the addition of saturated NaHCO₃ and extracted with DCM. Silica gelchromatography eluting with 0-5% MeOH:DCM yielded the title compound (22mg, 72%) as a cis/trans-mixture. ¹H NMR (CDCl₃, 300 MHz): δ 9.15 (d,J=4.8 Hz, 1H), 9.11 (bs, 1H), 9.02 (d, J=1.2 Hz, 1H), 8.98 (d, J=1.1,1H), 8.69 (dd, J=0.9, 2.9 Hz, 1H), 8.66 (dd, J=0.9, 2.8 Hz, 1H), 8.39(d, J=1.2 Hz, 1H), 8.37 (d, J=1.2 Hz, 1H), 7.95 (m, 1H), 7.93 (s, 1H),7.87 (d, J=1.1 Hz, 1H), 7.44 (m, 1H), 6.75 (s, 2H), 6.59 (s, 2H), 3.97(s, 6H), 3.91 (s, 3H), 3.90 (s, 6H), 3.86 (s, 3H).

Example 129 Preparation of (E)- and(Z)-3-pyridin-4-ylmethylene-5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one

(E)- and(Z)-3-pyridin-4-ylmethylene-5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-onewere prepared by a method analogous to that described in Example 128 bysubstituting 3-pyridinecarboxaldehyde for 4-pyridinecarboxaldehyde inthe reaction with Compound BN. The isomers were separated using silicagel chromatography eluting with 0-5% MeOH:DCM. Assignment ofstereochemistry is tentatively based on the ¹H NMR spectra. ¹H NMR(CDCl₃, 300 MHz): E-isomer: δ 8.91 (s, 1H), 8.76 (d, (J=3.6 Hz, 1H),8.39 (d, J=1.2 Hz, 1H), 8.02 (d, J=3.7 Hz, 1H), 7.91 (d, J=1.2 Hz, 1H),7.52 (s, 1H), 6.74 (s, 2H), 3.96 (s, 6H), 3.91 (s, 3H). Z-isomer: δ 9.01(s, 1H), 8.78 (d, (J=3.5 Hz, 1H), 8.38 (d, J=1.2 Hz, 1H), 7.87 (s, 1H),7.81 (d, J=1.2 Hz, 1H), 7.52 (d, J=6.1 Hz, 1H), 6.56 (s, 2H), 3.89 (s,6H), 3.88 (s, 3H).

Example 130 Preparation of3-benzylidene-5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one

3-Benzylidene-5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-onewas prepared by a method analogous to that described in Example 128 bysubstituting 3-pyridinecarboxaldehyde for benzaldehyde in the reactionwith Compound BN. 15 mg (33%) of the title compound were obtained.

Example 131 Preparation of4-[2-oxo-5-(3,4,5-trimethoxy-phenyl)-1,2-dihydro-pyrrolo[2,3-b]pyridin-3-ylidenemethyl]-benzamide

4-[2-Oxo-5-(3,4,5-trimethoxy-phenyl)-1,2-dihydro-pyrrolo[2,3-b]pyridin-3-ylidenemethyl]-benzamideonewas prepared by a method analogous to that described in Example 128 bysubstituting 3-pyridinecarboxaldehyde for 4-formylbenzamide in thereaction with Compound BN. 25 mg (50%) of the title compound wereobtained.

Example 132 Preparation of3-[2-oxo-5-(3,4,5-trimethoxy-phenyl)-1,2-dihydro-pyrrolo[2,3-b]pyridin-3-ylidenemethyl]-benzamide

3-[2-Oxo-5-(3,4,5-trimethoxy-phenyl)-1,2-dihydro-pyrrolo[2,3-b]pyridin-3-ylidenemethyl]-benzamideonewas prepared by a method analogous to that described in Example 128 bysubstituting 3-pyridinecarboxaldehyde for 3-formylbenzamide in thereaction with Compound BN. 26 mg (52%) of the title compound wereobtained.

Example 133

Preparation of5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one(Intermediate BY)

A mixture of 5-bromo-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one (200 mg,0.939 mmol), 3,4,5-trimethoxyphenylboronic acid (239 mg, 1.127 mmol) anddichlorobis(triphenylphosphine)palladium (II) (33 mg, 0.047 mmol) inCH₃CN (5 ml) and 1 M Na₂CO₃ (5 ml) was heated in a microwave reactor for10 min at 150° C. The reaction mixture was filtered, evaporated,partitioned between water and DCM and purified by silica gelchromatography with 0-10% MeOH:DCM to obtain 85 mg (30%) of compound #.¹H NMR (CDCl₃/DMSO-d₆, 300 MHz): δ 10.19 (bs, 1H), 8.18 (d, J=1.1 Hz,1H), 7.54 (s, 1H), 6.57 (s, 2H), 3.80 (s, 6H), 3.75 (s, 3H), 3.47 (s,2H).

Preparation of4-[2-oxo-5-(3,4,5-trimethoxy-phenyl)-1,2-dihydro-pyrrolo[2,3-b]pyridin-3-ylidenemethyl]-benzonitrile

A mixture of5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one(Intermediate BY, 42 mg, 0.14 mmol), 4-cyanobenzaldehyde (22 mg, 0.168mmol), triethylamine (22 μl, 0.168 mmol) and molecular sieves 4 Å (100mg) in toluene (2 ml) was reacted at 80° C. for 1 d. The mixture waspartitioned between DCM and water, the aqueous phase extracted with DCM,combined organic phases dried, evaporated and purified by silica gelchromatography (0-5% MeOH:DCM) to obtain 31 mg (54%) of the titlecompound as a mixture of (E)- and (Z)-isomers.

Example 134 Preparation of3-[2-oxo-5-(3,4,5-trimethoxy-phenyl)-1,2-dihydro-pyrrolo[2,3-b]pyridin-3-ylidenemethyl]-benzonitrile

3-[2-oxo-5-(3,4,5-trimethoxy-phenyl)-1,2-dihydro-pyrrolo[2,3-b]pyridin-3-ylidenemethyl]-benzonitrilewas prepared by a method analogous to that described in Example 133 bysubstituting 4-cyanobenzaldehyde for 3-cyanobenzaldehyde in the reactionwith Intermediate BY. 36 mg (62%) of the title compound were obtained asa mixture of cis- and trans-isomers.

Example 135

Preparation of3-pyridin-3-ylmethyl-5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one

To a solution of3-pyridin-4-ylmethylene-5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one(50 mg, 0.128 mmol) in MeOH (4 ml) was added ammonium formate (245 mg,3.85 mmol) and Pd/C (10%, 30 mg). The mixture was stirred at roomtemperature for 3 hrs after which it was filtered, evaporated, andpartitioned between water and DCM. The title compound (33 mg, 66%) wasobtained after silica gel chromatography eluting with 0-10% MeOH:DCM. ¹HNMR (CDCl₃, 300 MHz): δ 10.05 (s, 1H), 8.60 (d, J=2.6 Hz, 1H), 8.45 (d,J=1.1 Hz, 1H), 8.38 (d, J=1.2 Hz, 1H), 7.62 (d, J=4.7 Hz, 1H), 7.35 (dd,J=2.9, 4.7 Hz, 1H), 6.53 (d, J=1.2 Hz, 1H), 6.38 (s, 1H), 3.95 (m, 1H),3.90 (m, 1H), 3.85 (s, 6H), 3.84 (s, 3H), 3.84 (m, 1H).

Example 136 Preparation of3-pyridin-4-ylmethyl-5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one

3-Pyridin-4-ylmethyl-5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-onewas prepared by a method analogous to that described in Example 135. Thetitle compound (14 mg, 61%) was obtained after silica gel chromatographyeluting with 0-8% MeOH:DCM. ¹H NMR (CDCl₃, 300 MHz): δ 9.52 (bs, 1H),8.54 (d, J=3.5 Hz, 1H), 8.32 (d, J=1.1 Hz, 1H), 7.18 (d, J=3.6 Hz, 1H),7.12 (m, 1H), 6.54 (s, 1H), 3.91 (s, 6H), 3.89 (m, 1H), 3.88 (s, 3H),3.54 (dd, J=3.1, 8.3 Hz, 1H), 3.03 (dd, J=5.6, 8.3 Hz, 1H).

Example 137

Preparation of3-benzyl-5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one

To a solution of3-benzylidene-5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one(41 mg, 0.106 mmol) in a mixture of MeOH (2 ml), THF (1 ml) and water(0.3 ml) was added sodium borohydride (40 mg, 1.06 mmol). The reactionwas stirred at room temperature for 10 min after which it was quenchedby the addition of 1 N HCl and partitioned between water and DCM. Theresidue was purified by preparatory HPLC to yield the title compound(5.2 mg, 13%).

Example 138 Preparation of4-[2-oxo-5-(3,4,5-trimethoxy-phenyl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl]-benzamide

4-[2-Oxo-5-(3,4,5-trimethoxy-phenyl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl]-benzamidewas prepared from4-[2-oxo-5-(3,4,5-trimethoxy-phenyl)-1,2-dihydro-pyrrolo[2,3-b]pyridin-3-ylidenemethyl]-benzamideoneby a method analogous to that described in Example 137. The titlecompound (12 mg, 54%) was obtained after silica gel chromatographyeluting with 0-10% MeOH:DCM. ¹H NMR (DMSO-d6, 300 MHz): δ 11.06 (s, 1H),8.34 (d, J=1.5 Hz, 1H), 7.88 (s, 1H), 7.75 (d, J=5.0 Hz, 2H), 7.41 (d,J=0.5 Hz, 1H), 7.30 (s, 1H), 7.28 (d, J=5.0 Hz, 2H), 6.74 (s, 2H), 4.03(m, 1H), 3.82 (s, 6H), 3.67 (s, 3H), 3.44 (dd, J=3.4, 8.2 Hz, 1H), 3.11(dd, J=4.6, 8.2 Hz, 1H).

Example 139 Preparation of3,3-dibenzyl-5-(3,4,5-trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one

5-(3,4,5-Trimethoxy-phenyl)-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one (95mg, 0.316 mmol) and TMEDA (96 μl, 0.623 mmol) were dissolved inanhydrous THF (4 ml) and cooled to −78° C. n-BuLi (1.6 M in hexanes, 415μl, 0.664 mmol) was added dropwise. After completed addition stiffingwas continued for 1 hr at −78° C. Benzyl bromide (41.3 μl, 0.348 mmol)was added dropwise as a 10% solution in anh. THF. After completedaddition the reaction was allowed to warm up to room temperature whilestiffing overnight. The reaction was quenched by the addition of MeOH,evaporated and partitioned between water and DCM. Silica gelchromatography eluting with 0-50% EtOAc:Hexanes yielded the titlecompound (47 mg, 38%). ¹H NMR (CDCl₃, 300 MHz): δ 8.83 (s, 1H), 8.18 (d,J=1.2 Hz, 1H), 7.19 (d, J=1.2 Hz, 1H), 7.14 (m, 6H), 6.99 (m, 4H), 6.61(s, 2H), 3.96 (s, 6H), 3.90 (s, 3H), 3.30 (d, J=8.0 Hz, 2H), 3.26 (d,J=8.0 Hz, 2H).

Example 140 Preparation of1-(4-{4-[3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzyl}-piperazin-1-yl)-ethanone

2-(4-Bromomethyl-phenyl)-4,4,5,5-tetramethyl-[1,3,2]dioxaborolane (100mg, 0.337 mmol), N-acetylpiperazine (47 mg, 0.37 mmol) and K₂CO₃ (93 mg,0.675 mmol) were combined in DMF (2.5 ml) and stirred overnight at roomtemperature. The reaction was quenched by the addition of water,extracted with DCM and dried. The residue was taken up in CH₃CN (2 ml),Intermediate B (120 mg, 0.275 mmol) anddichlorobis(triphenylphosphine)palladium (II) (10 mg, 0.013 mmol) wereadded and the reaction was heated to 150° C. in a microwave reactor for20 min. The mixture was partitioned between water and DCM, the organicphase dried, evaporated and purified by silica gel chromatography using0-5% MeOH:DCM. 53 mg (46%) of the title compound were obtained. MS ESI(m/z): 450.4 (M+1)⁺, calc. 449.

Example 141 Preparation of4-{4-[3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzyl}-1-methyl-piperazin-2-one

4-{4-[3-(1H-Indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzyl}-1-methyl-piperazin-2-onewas prepared by a method analogous to that described in Example 140 bysubstituting N-acetylpiperazine for 1-methyl-piperazin-2-one. The titlecompound (14 mg, 28%) was obtained after silica gel chromatographyeluting with 0-10% MeOH:DCM. MS ESI (m/z): 435.9 (M+1)⁺, calc. 435.

Example 142 Preparation of4-{4-[3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzyl}-piperazin-2-one

4-{4-[3-(1H-Indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzyl}-piperazin-2-onewas prepared by a method analogous to that described in Example 140 bysubstituting N-acetylpiperazine for piperazin-2-one. The title compound(22 mg, 45%) was obtained after silica gel chromatography eluting with0-10% MeOH:DCM. MS ESI (m/z): 422.2 (M+1)⁺, calc. 421.

Example 143 Preparation of4-{3-[3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzyl}-1-methyl-piperazin-2-one

4-{3-[3-(1H-Indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzyl}-1-methyl-piperazin-2-onewas prepared by a method analogous to that described in Example 140 bysubstituting N-acetylpiperazine for 1-methyl-piperazin-2-one and2-(4-bromomethyl-phenyl)-4,4,5,5-tetramethyl-[1,3,2]dioxaborolane for3-(bromomethyl)phenylboronic acid. The title compound (22 mg, 45%) wasobtained after silica gel chromatography eluting with 0-10% MeOH:DCM. MSESI (m/z): 436.4 (M+1)⁺, calc. 435.

Example 144 Preparation of4-{4-[3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzyl}-piperazine-1-carboxylicacid tert-butyl ester

4-{4-[3-(1H-Indol-5-3/0-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzyl}-piperazine-1-carboxylicacid tert-butyl ester was prepared by a method analogous to thatdescribed in Example 140 by substituting N-acetylpiperazine forN-Boc-piperazine. The title compound (20 mg, 33%) was obtained aftersilica gel chromatography eluting with 0-3% MeOH:DCM. MS ESI (m/z):508.2 (M+1)⁺, calc. 507.

Example 145

Preparation of{4-[3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-phenyl}-(4-methyl-piperazin-1-yl)-methanone

4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzoic acid (100 mg,0.403 mmol), EDCI (97 mg, 0.504 mmol) and DMAP (catalytic amount) werecombined in CH₃CN, stirred for 10 min and treated withN-methylpiperazine (54 μl, 0.484 mmol). The mixture was stirredovernight at room temperature. An aliquot of 650 μl was taken, combinedwith Intermediate B (50 mg, 0.107 mmol) anddichlorobis(triphenylphosphine)palladium (II) (10 mg, 0.013 mmol) andheated to 150° C. in a microwave reactor for 20 min. The mixture waspartitioned between water and DCM, the organic phase dried, evaporatedand purified by silica gel chromatography using 0-6% MeOH:DCM. 13 mg(28%) of the title compound were obtained. ¹H NMR (DMSO-d6, 300 MHz): δ11.88 (d, J=1.5 Hz, 1H), 11.08 (s, 1H), 8.57 (d, J=2.1 Hz, 1H), 8.45 (d,J=1.8 Hz, 1H), 7.90 (s, 1H), 7.82 (d, J=8.4 Hz, 1H), 7.77 (d, J=2.4 Hz,1H), 7.47 (m, 4H), 7.34 (t, J=2.6 Hz, 1H), 6.47 (t, J=2.4 Hz, 1H), 3.58(bs, 4H), 2.3 (bs, 4H), 2.18 (s, 3H).

Example 146 Preparation of1-(4-{4-[3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzoyl}-piperazin-1-yl)-ethanone

1-(4-{4-[3-(1H-Indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzoyl}-piperazin-1-yl)-ethanonewas synthesized by a method analogous to that described in Example 144by substituting N-methylpiperazine for N-acetylpiperazine. The titlecompound (13 mg, 26%) was obtained after silica gel chromatographyeluting with 0-5% MeOH:DCM. MS ESI (m/z): 464.2 (M+1)⁺, calc. 463.

Example 147 Preparation of{3-[3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-phenyl}-(4-methyl-piperazin-1-yl)-methanone

{3-[3-(1H-Indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-phenyl}-(4-methyl-piperazin-1-yl)-methanonewas synthesized by a method analogous to that described in Example 144by substituting 4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzoicacid for 3-carboxyphenylboronic acid. The title compound (23 mg, 49%)was obtained after silica gel chromatography eluting with 5-10%MeOH:DCM. MS ESI (m/z): 436.4 (M+1)⁺, calc. 435.

Example 148

Preparation of4-[5-bromo-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzamide(Intermediate CA)

5-Bromo-3-iodo-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridine(Intermediate BZ, 483 mg, 1.01 mmol), 4-aminocarbonylphenylboronic acid(196 mg, 1.22 mmol) and dichlorobis(triphenylphosphine)palladium (II)(71 mg, 0.1 mmol) were combined in CH₃CN (10 ml) and 1 M Na₂CO₃ (10 ml)and stirred at 60° C. for 3 hrs. Water was added and the mixture wasextracted with DCM and purified by silica gel chromatography using 0-30%EtOAc/Hexanes. The title compound was obtained in 79% yield (373 mg). ¹HNMR (CDCl₃, 300 MHz): δ 8.51 (d, J=1.2 Hz, 1H), 8.20 (d, J=1.2 Hz, 1H),8.11 (d, J=5.1 Hz, 2H), 7.96 (s, 1H), 7.93 (d, J=5.0 Hz, 2H), 7.64 (d,J=5.1 Hz, 2H), 7.31 (d, J=4.8 Hz, 2H), 6.1 (bs, 1H), 5.7 (bs, 1H), 2.39(s, 3H).

Preparation of4-{4-[3-(4-carbamoyl-phenyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzoyl}-piperazine-1-carboxylicacid tert-butyl ester

4-[5-Bromo-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzamide(Intermediate CA, 200 mg, 0.425 mmol),4-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzoyl]-piperazine-1-carboxylicacid tert-butyl ester (212 mmg, 0.51 mmol) anddichlorobis(triphenylphosphine)palladium (II) (15 mg, 0.021 mmol) werecombined in CH₃CN (5 ml) and 1 M Na₂CO₃ (5 ml) and reacted in amicrowave reactor at 150° C. for 10 min. The mixture was filtered, waterwas added, extracted with EtOAc and purified by silica gelchromatography using 0-8% MeOH:DCM. The title compound was obtained in46% yield (102 mg). ¹H NMR (DMSO-d6, 300 MHz): δ 12.2 (bs, 1H), 8.63 (d,J=1.1 Hz, 1H), 8.54 (d, J=1.1 Hz, 1H), 8.08 (s, 1H), 7.98 (bs, 1H), 7.96(d, J=5.1 Hz, 2H), 7.89 (m, 4H), 7.54 (d, J=4.9 Hz, 2H), 7.32 (bs, 1H),3.6 (bs, 2H), 3.4 (bs), 1.41 (s, 9H).

Example 149 Preparation of4-{5-[4-(piperazine-1-carbonyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamide,hydrochloride salt

A solution of4-{4-[3-(4-carbamoyl-phenyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzoyl}-piperazine-1-carboxylicacid tert-butyl ester (100 mg, 0.19 mmol) in MeOH (3 ml) was treatedwith 4 N HCl in dioxane (2.5 ml) and stirred at room temperature for 1hr. The mixture was evaporated, taken up in MeOH and evaporated again.This was repeated twice to give 102 mg (116%) of the title compound. MSESI (m/z): MS ESI (m/z): 426.4 (M+1)⁺, calc. 425.

Example 150 Preparation of4-{5-[4-(4-acetyl-piperazine-1-carbonyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamide

To a solution of4-{5-[4-(piperazine-1-carbonyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamide,hydrochloride salt (19 mg, 0.041 mmol) in MeOH (2 ml) was addedtriethylamine (400 μl, 2.88 mmol) and acetic anhydride (100 μl, 1.06mmol). The mixture was stirred for 1 hr at room temperature. EtOAc wasadded and washed with saturated aqu. NaHCO₃, water, brine and dried andevacuated. Purification on silica gel employing 0-10% MeOH:DCM provided4.7 mg (25%) of the title compound. MS ESI (m/z): 468.3 (M+1)⁺, calc.467.

Example 151 Preparation of4-{3-[3-(4-carbamoyl-phenyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzoyl}-piperazine-1-carboxylicacid tert-butyl ester

4-{3-[3-(4-Carbamoyl-phenyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzoyl}-piperazine-1-carboxylicacid tert-butyl ester was prepared by a method analogous to thatdescribed in Example 148 by substituting4-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzoyl]-piperazine-1-carboxylicacid tert-butyl ester for4-[3-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzoyl]-piperazine-1-carboxylicacid tert-butyl ester. The title compound (109 mg, 49%) was obtainedafter silica gel chromatography eluting with 0-8% MeOH:DCM. ¹H NMR(DMSO-d6, 300 MHz): δ 12.18 (bs, 1H), 8.61 (d, J=1.2 Hz, 1H), 8.52 (d,J=1.2 Hz, 1H), 8.07 (s, 1H), 7.96 (m, 3H), 7.89 (m, 3H), 7.80 (s, 1H),7.57 (t, J=4.6 Hz, 1H), 7.41 (d, J=4.6 Hz, 1H), 7.32 (s, 1H), 3.63 (bs,2H), 3.4 (bs, 2H), 1.40 (s, 9H).

Example 152 Preparation of4-{5-[3-(piperazine-1-carbonyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamide,hydrochloride salt

The hydrochloride salt of4-{5-[3-(piperazine-1-carbonyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamidewas prepared by a method analogous to that described in Example 149 bysubstituting4-{4-[3-(4-carbamoyl-phenyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzoyl}-piperazine-1-carboxylicacid tert-butyl ester for4-{3-[3-(4-carbamoyl-phenyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzoyl}-piperazine-1-carboxylicacid tert-butyl ester. 105 mg (128%) of the title compound wereobtained. ¹H NMR (DMSO-d6, 300 MHz): δ 12.32 (s, 1H), 9.52 (s, 2H), 8.66(d, J=1.8 Hz, 1H), 8.59 (d, J=1.8 Hz, 1H), 8.11 (d, J=2.7 Hz, 1H), 7.95(m, 5H), 7.60 (t, J=7.8 Hz, 1H), 7.52 (d, J=7.2 Hz, 1H), 7.36 (bs, 1H),3.6-4.0 (bs, 8H).

Example 153 Preparation of4-{5-[3-(4-acetyl-piperazine-1-carbonyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamide

4-{5-[3-(4-Acetyl-piperazine-1-carbonyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamidewas prepared by a method analogous to that described in Example 150 bysubstituting4-{5-[4-(piperazine-1-carbonyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamide,hydrochloride salt for4-{5-[3-(piperazine-1-carbonyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamide,hydrochloride salt. 3.1 mg (14%) of the title compound were obtained. ¹HNMR (CD₃OD, 300 MHz): δ 8.56 (d, J=1.2 Hz, 1H), 8.55 (d, J=1.2 Hz, 1H),7.99 (d, J=4.2 Hz, 2H), 7.86 (m, 4H), 7.81 (d, J=1.8 Hz, 1H), 7.62 (t,J=4.6 Hz, 1H), 7.47 (dd, J=0.7, 3.8 Hz, 1H), 3.5-3.9 (m, 8H), 2.14 (bd,3H).

Example 154 Preparation of4-[7-oxy-5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzamide

4-[5-(3,4,5-Trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzamide(50 mg, 0.124 mmol), magnesium monoperoxyphthalic acid (80%, 300 mg,0.46 mmol) and acetic acid (10 drops were combined in EtOH (3 ml) andstirred at 50° C. for 1 hr. After adding EtOAc the mixture was washedwith saturated NaHCO₃, dried and purified by silica gel chromatographyusing 0-8% MeOH:DCM to provide 18 mg (33%) of the title compound. ¹H NMR(DMSO-d6, 300 MHz): δ 12.9 (bs, 1H), 8.62 (s, 1H), 8.14 (s, 1H), 8.0(bs, 2H), 7.97 (d, J=5.0 Hz, 2H), 7.89 (d, J=5.0 Hz, 2H), 7.34 (bs, 1H),7.04 (s, 2H), 3.89 (s, 6H), 3.70 (s, 3H).

Example 155 Preparation of4-{5-[4-(4-methyl-piperazin-1-ylmethyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamide

4-{5-[4-(4-Methyl-piperazin-1-ylmethyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamidewas prepared by a method analogous to that described in Example 148 bysubstituting4-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzoyl]-piperazine-1-carboxylicacid tert-butyl ester for1-methyl-4-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzyl]-piperazine.The title compound (24 mg, 44%) was obtained by precipitation from DCM.¹H NMR (DMSO-d6, 300 MHz): δ 12.1 (s, 1H), 8.57 (d, J=1.2 Hz, 1H), 8.48(d, J=1.2 Hz, 1H), 8.05 (d, J=1.3 Hz, 1H), 7.98 (bs, 1H), 7.96 (d, J=5.0Hz, 2H), 7.88 (d, J=5.1 Hz, 2H), 7.73 (d, J=4.5 Hz, 2H), 7.40 (d, J=4.5Hz, 2H), 7.31 (bs, 1H), 3.50 (s, 2H), 2.2-2.45 (bs, 8H), 2.15 (s, 3H).

Example 156 Preparation of4-{5-[3-(4-methyl-piperazin-1-ylmethyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamide

4-{5-[3-(4-Methyl-piperazin-1-ylmethyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamidewas prepared by a method analogous to that described in Example 148 bysubstituting4-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzoyl]-piperazine-1-carboxylicacid tert-butyl ester for1-methyl-4-[3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzyl]-piperazine.The title compound (8 mg, 15%) was obtained by precipitation from DCM.¹H NMR (DMSO-d6, 300 MHz): δ 12.1 (s, 1H), 8.56 (d, J=1.2 Hz, 1H), 8.46(d, J=1.2 Hz, 1H), 8.05 (d, J=1.3 Hz, 1H), 7.96 (m, 3H), 7.88 (d, J=5.1Hz, 2H), 7.66 (m, 2H), 7.45 (m, 1H), 7.31 (m, 2H), 3.55 (s, 2H),2.2-2.45 (bs, 8H), 2.14 (s, 3H).

Example 157 Preparation of4-{5-[4-(4-acetyl-piperazin-1-ylmethyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamide

4-{5-[4-(4-Acetyl-piperazin-1-ylmethyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamidewas prepared by a method analogous to that described in Example 140 bysubstituting Intermediate B with Intermediate CA. Purification by silicagel chromatography using 4-5% MeOH:DCM yielded the title compound (13mg, 30%). MS ESI (m/z): 454.1 (M+1)⁺, calc. 453.

Example 158 Preparation of4-{5-[4-(4-methyl-3-oxo-piperazin-1-ylmethyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamide

4-{5-[4-(4-Methyl-3-oxo-piperazin-1-ylmethyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamidewas prepared by a method analogous to that described in Example 140 bysubstituting Intermediate B with Intermediate CA and N-acetylpiperazinefor 1-methyl-piperazin-2-one. Purification by silica gel chromatographyusing 4-5% MeOH:DCM yielded the title compound (4 mg, 10%). MS ESI(m/z): 440.3 (M+1)⁺, calc. 439.

Example 159

Preparation of4-[5-bromo-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzylamine(Intermediate CB)

5-Bromo-3-iodo-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridine (200 mg,0.419 mmol), 4-aminomethylphenylboronic acid hydrochloride (95 mg, 0.503mmol) and dichlorobis(triphenylphosphine)palladium (II) (29 mg, 0.042mmol) were combined in CH₃CN (5 ml) and 1 M Na₂CO₃ (5 ml) and stirred at60° C. for 3 hrs. EtOAc was added, the organic phase was washed withwater, dried and evaporated. to yield 136 mg (71%) of the titlecompound. MS ESI (m/z): 455.9/458.1 (M+1)⁺, calc. 455/457.

Preparation ofN-{4-[5-bromo-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzyl}-acetamide(Intermediate CC)

4-[5-Bromo-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzylamine(Intermediate CB, 45 mg, 0.1 mmol) was combined with triethylamine (45μl, 0.3 mmol) and acetic anhydride (11 μl, 0.11 mmol) in anh. DCM (2ml). The mixture was stirred for 2 hrs, EtOAc, was added and washed with0.5 N HCl, saturated NaHCO₃, water and brine. Evaporation yielded thetitle compound (48 mg, 96%). MS ESI (m/z): 498.1/500.1 (M+1)⁺, calc.497/499.

Preparation ofN-{4-[5-(3,4,5-Trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzyl}-acetamide

N-{-4-[5-Bromo-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzyl}-acetamide(Intermediate CC, 24 mg, 0.048 mmol), 3,4,5-trimethoxyphenyl boronicacid (13 mg, 0.058 mmol) and dichlorobis(triphenylphosphine)palladium(II) (2 mg, 0.002 mmol) were combined in CH₃CN (1 ml) and 1 M Na₂CO₃ (2ml) and heated in a microwave reactor at 150° C. for 20 min. EtOAc wasadded, washed with water, dried and purified by silica gelchromatography eluting with 0-4% MeOH:DCM to give 11 mg (53%) of thetitle compound. MS ESI (m/z): 432.2 (M+1)⁺, calc. 431.

Example 160 Preparation of2-phenyl-N-{4-[5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzyl}-acetamide

2-Phenyl-N-{4-[5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzyl}-acetamidewas prepared by a method analogous to that described in Example 159 bysubstituting acetic anhydride for phenacetyl chloride. Purification bysilica gel chromatography using 0-4% MeOH:DCM yielded the title compound(9 mg, 38%). MS ESI (m/z): 508.3 (M+1)⁺, calc. 507.

Example 161 Preparation of3-phenyl-N-{4-[5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzyl}-propionamide

3-Phenyl-N-{4-[5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzyl}-propionamidewas prepared by a method analogous to that described in Example 159 bysubstituting acetic anhydride for phenylpropionyl chloride. Purificationby silica gel chromatography using 0-4% MeOH:DCM yielded the titlecompound (13 mg, 54%). MS ESI (m/z): 522.4 (M+1)⁺, calc. 521.

Example 162

Preparation of5-bromo-3-[4-(4-methyl-piperazin-1-ylmethyl)-phenyl]-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridine(Intermediate CD)

5-Bromo-3-iodo-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridine (200 mg,0.419 mmol),1-methyl-4-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzyl]-piperazine(160 mg, 0.503 mmol) and dichlorobis(triphenylphosphine)palladium (II)(30 mg, 0.042 mmol) were combined in CH₃CN (5 ml) and 1 M Na₂CO₃ (5 ml)and stirred at 60° C. for 2 hrs. EtOAc was added and the organic phasewas washed with water, dried and evaporated. Purification by silica gelchromatography using 0-20% MeOH:DCM yielded 235 mg (104%) of the titlecompound. MS ESI (m/z): 539.0/541.2 (M+1)⁺, calc. 538/540.

Preparation of4-{3-[4-(4-methyl-piperazin-1-ylmethyl)-phenyl]-1,1-pyrrolo[2,3-b]pyridin-5-yl}-benzamide

5-Bromo-3-[4-(4-methyl-piperazin-1-ylmethyl)-phenyl]-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridine(Intermediate CD, 70 mg, 0.13 mmol), aminocarbonylphenylboronic acid (26mg, 0.156 mmol) and dichlorobis(triphenylphosphine)palladium (II) (5 mg,0.0065 mmol) were combined in CH₃CN (2 ml) and 1 M Na₂CO₃ (2 ml) andreacted in a microwave reactor for 20 min at 150° C. Water was added andthe aqueous phase was extracted with DCM, dried and evaporated.Purification by reversed phase chromatography using 0-100% MeOH:wateryielded 6 mg (11%) of the title compound. MS ESI (m/z): 426.7 (M+1)⁺,calc. 425.

Example 163 Preparation of5-(1H-indol-5-yl)-3-[4-(4-methyl-piperazin-1-ylmethyl)-phenyl]-1H-pyrrolo[2,3-b]pyridine

5-(1H-Indol-5-yl)-3-[4-(4-methyl-piperazin-1-ylmethyl)-phenyl]-1H-pyrrolo[2,3-b]pyridinewas prepared by a method analogous to that described in Example 162 bysubstituting aminocarbonylphenylboronic acid for indole-5-boronic acid.Purification by silica gel chromatography using 0-10% MeOH:DCM yieldedthe title compound (28 mg, 60%). MS ESI (m/z): 422.4 (M+1)⁺, calc. 421.

Example 164

Preparation of4-[5-bromo-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-methyl-benzamide(Intermediate CE)

5-Bromo-3-iodo-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridine (350 mg,0.73 mmol), 4-(N-methylaminocarbonyl)phenylboronic acid (160 mg, 0.88mmol) and dichlorobis(triphenylphosphine)palladium (II) (52 mg, 0.073mmol) were combined in CH₃CN (10 ml) and 1 M Na₂CO₃ (10 ml) and stirredat 60° C. for 5 hrs. Water was added and the mixture was extracted withDCM, combined organic phases were dried and evaporated to yield 428 mg(121%) of the title compound. ¹H NMR (CDCl₃, 300 MHz): δ 8.50 (d, J=1.3Hz, 1H), 8.20 (d, J=1.2 Hz, 1H), 8.09 (d, J=5.1 Hz, 2H), 7.94 (s, 1H),7.87 (d, J=5.1, 2H), 7.61 (d, J=5.0 Hz, 2H), 7.31 (d, J=5.0 Hz, 2H),6.21 (bd, J=2.5 Hz, 1H), 3.06, (d, J=2.9 Hz, 3H), 2.39 (s, 3H).

Preparation ofN-methyl-4-[5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-benzamide

4-[5-Bromo-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-methyl-benzamide(Intermediate CE, 100 mg, 0.206 mmol), 3,4,5-trimethoxyphenylboronicacid (53 mg, 0.248 mmol) and dichlorobis(triphenylphosphine)palladium(II) (9 mg, 0.012 mmol) were combined in CH₃CN (2 ml) and 1 M Na₂CO₃ (2ml) and reacted in a microwave reactor for 20 min at 150° C. Water wasadded, the aqueous phase was extracted with DCM and the organic phasewas dried and evaporated. Purification by silica gel chromatographyusing 0-8% MeOH:DCM yielded 40 mg (47%) of the title compound. ¹H NMR(CDCl₃, 300 MHz): δ 12.09 (s, 1H), 8.59 (d, J=1.2 Hz, 1H), 8.48 (d,J=1.2 Hz, 1H), 8.43 (q, J=2.7 Hz, 1H), 8.04 (s, 1H), 7.94 (d, J=4.0,2H), 7.91 (d, J=4.0 Hz, 2H), 7.00 (s, 2H), 3.89 (s, 6H), 3.70 (s, 3H),2.80, (d, J=4.5 Hz, 3H).

Example 165 Preparation ofN-methyl-4-{5-[4-(4-methyl-piperazin-1-ylmethyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamide

N-Methyl-4-{5-[4-(4-methyl-piperazin-1-ylmethyl)-phenyl]-1H-pyrrolo[2,3-b]pyridin-3-yl}-benzamidewas prepared by a method analogous to that described in Example 164 bysubstituting 3,4,5-trimethoxyphenylboronic acid for1-methyl-4-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzyl]-piperazine.Purification by precipitation from hot DCM yielded the title compound(46 mg, 51%). ¹H NMR (CDCl₃, 300 MHz): δ 12.09 (s, 1H), 8.57 (d, J=1.2Hz, 1H), 8.48 (d, J=1.2 Hz, 1H), 8.43 (q, J=2.7 Hz, 1H), 8.05 (d, J=1.5Hz, 1H), 7.92 (d, J=5.2 Hz, 2H), 7.89 (d, J=5.2 Hz, 2H), 7.73 (d, J=4.9Hz, 2H), 7.40 (d, J=4.9 Hz, 2H), 3.50 (s, 2H), 2.81, (d, J=2.7 Hz, 3H),2.2-2.45 (bs. 8H), 2.15 (s, 3H).

Example 166

Preparation of5-bromo-3-(4-fluoro-phenyl)-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridine(Intermediate CF)

5-Bromo-3-iodo-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridine (70 mg,0.147 mmol), 4-fluorophenylboronic acid (25 mg, 0.176 mmol) anddichlorobis(triphenylphosphine)palladium (II) (10 mg, 0.015 mmol) werecombined in CH₃CN (2 ml) and 1 M Na₂CO₃ (2 ml) and stirred at 60° C. for3 hrs. EtOAc was added and the mixture was washed with water, dried andevaporated to yield 73 mg (112%) of the title compound. MS ESI (m/z):445.1/447.2 (M+1)⁺, calc. 444/446.

Preparation of3-(4-fluoro-phenyl)-5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine

5-Bromo-3-(4-fluoro-phenyl)-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridine(37 mg, 0.083 mmol), 3,4,5-trimethoxyphenylboronic acid (21 mg, 0.1mmol) and dichlorobis(triphenylphosphine)palladium (II) (3 mg, 0.004mmol) were combined in CH₃CN (1.5 ml) and 1 M Na₂CO₃ (2 ml) and reactedin a microwave reactor for 20 min at 150° C. EtOAc was added and themixture was washed with water, dried, evaporated and purified by silicagel chromatography using 0-2% MeOH:DCM to yield 9 mg (29%) of the titlecompound. MS ESI (m/z): 379.2 (M+1)⁺, calc. 378.

Example 167

Preparation of2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-1H-imidazole(Intermediate CG)

2-(4-Bromo-phenyl)-1H-imidazole (300 mg, 1.3 mmol),bis(pinacolato)diboron (376 mg, 1.48 mmol), KOAc (400 mg, 4.03 mmol) andPdCl₂(dppf) CH₂Cl₂ (50 mg, 0.067 mmol) were combined in DMSO (8 ml) andstirred t 80° C. overnight. EtOAc was added, washed with water, dried,evaporated and purified by silica gel chromatography eluting with 0-5%MeOH:DCM to give 116 mg (36%) of the title compound. ¹H NMR (CDCl₃, 300MHz): δ 7.86 (s, 4H), 7.18 (s, 2H), 1.36 (s, 12H).

Preparation of 5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine(Intermediate CH)

5-Bromo-1H-pyrrolo[2,3-b]pyridine (1.54 g, 7.83 mmol),3,4,5-trimethoxyphenylboronic acid (1.83 g, 8.61 mmol) anddichlorobis(triphenylphosphine)palladium (II) (275 mg, 0.39 mmol) werecombined in CH₃CN (10 ml) and 1 M Na₂CO₃ (10 ml) and reacted in amicrowave reactor for 5 min at 150° C. EtOAc was added and the mixturewas washed with water, brine, dried, evaporated and purified by silicagel chromatography using 0-2% MeOH:DCM to yield 1.86 g (84%) of thetitle compound. ¹H NMR (CDCl₃, 300 MHz): δ 9.9 (bs, 1H), 8.54 (d, J=2.1Hz, 1H), 8.11 (d. J=2.1 Hz, 1H), 7.41 (t, J=2.1 Hz, 1H), 6.82 (s, 2H),6.58 (t, J=1.5 Hz, 1H), 3.96 (s, 6H), 3.92 (s, 3H).

Preparation of3-iodo-5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine(Intermediate CI)

To a solution of 5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine(510 mg, 1.79 mmol) in acetone (100 ml) was added N-iodosuccinimide (444mg, 1.97 mmol) under stiffing. After 1 hr the mixture was evaporated andpurified by silica gel chromatography using 0-2% MeOH:DCM to give thetitle compound (870 mg, 118%). MS ESI (m/z): 411.1 (M+1)⁺, calc. 410.

Preparation of3-iodo-1-(toluene-4-sulfonyl)-5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine(Intermediate CJ)

A solution of3-iodo-5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine (870 mg,2.12 mmol) in anh. THF (10 ml) was cooled to 0° C. and NaH (60%dispersion, 130 mg, 3.18 mmol) was added. After 20 min tosyl chloride(450 mg, 2.33 mmol) was added and the mixture was allowed to warm toroom temperature. After 3 hrs the mixture was cooled to 0° C. andquenched by the addition of 0.5 N HCl. The product was extracted withDCM and purified by silica gel chromatography using DCM as an eluentaffording 648 mg (54%). ¹H NMR (CDCl₃, 300 MHz): δ 8.61 (d, J=2.4 Hz,1H), 8.12 (d. J=8.4 Hz, 1H), 7.91 (s, 1H), 7.74 (d, J=2.1 Hz, 1H), 7.31(d, J=8.4 Hz, 2H), 6.73 (s, 2H), 3.94 (s, 6H), 3.90 (s, 3H), 2.39 (s,3H).

Preparation of3-[4-(1H-imidazol-2-yl)-phenyl]-5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine

3-Iodo-1-(toluene-4-sulfonyl)-5-(3,4,5-trimethoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine(Intermediate CJ, 30 mg, 0.053 mmol),2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-1H-imidazole(Intermediate CG, 18 mg, 0.064 mmol) anddichlorobis(triphenylphosphine)palladium (II) (2 mg, 0.003 mmol) werecombined in CH₃CN (1 ml) and 1 M Na₂CO₃ (1 ml) and stirred at 60° C. for2 d. Additional Intermediate CG (18 mg, 0.064 mmol) was added andstirring was continued for another day. EtOAc was added and the mixturewas washed with water, dried, evaporated and purified by silica gelchromatography using 0-5% MeOH:DCM to yield 5 mg (22%) of the titlecompound. MS ESI (m/z): 427.2 (M+1)⁺, calc. 426.

Biological Activity

The activity of the compounds in Examples 1-138 as MLK inhibitors isillustrated in the following assays. The other compounds listed above,which have not yet been made and/or tested, are predicted to haveactivity in these assays as well.

Radiometric filter plate MLK3 assay

200 ng (130 nM) MLK3 (Dundee, DU8313) was incubated with 1 μM inactiveMKK7b (Dundee, DU703) in the presence of 2 μM cold ATP (K_(m)) and 0.5μCi/assay ³³P ATP and appropriate concentrations of compounds. After atwenty minute incubation, the reactions were washed through filterplates and read on a scintillation counter. Results are shown in Table 4below, in which +++ indicates ≦0.1 μM, ++ indicates >0.1 μM and ≦1 μM,and + indicates >1 μM.

TABLE 4 Ex. MK3 IC₅₀ MW 1 ++ 399.45 2 ++ 369.43 3 +++ 366.50 4 +++325.50 5 +++ 354.50 6 ++ 340.39 7 +++ 408.50 8 +++ 324.50 9 +++ 326.3610 ++ 394.48 11 +++ 402.50 12 ++ 348.41 13 ++ 353.43 14 ++ 309.13 15 +++375.43 16 +++ 417.47 17 +++ 377.41 18 +++ 376.42 19 +++ 404.47 20 +304.32 21 ++ 358.45 22 + 366.83 23 ++ 301.35 24 +++ 413.48 25 +++ 403.4426 +++ 373.14 27 ++ 358.14 28 ++ 329.13 29 + 338.77 30 +++ 339.15 31 ++422.54 32 +++ 421.55 33 +++ 366.47 34 +++ 467.58 35 + 426.52 36 +++400.44 37 +++ 370.41 38 +++ 355.40 39 +++ 469.55 40 +++ 417.43 41 ++431.16 42 + 357.12 43 + 369.16 44 + 328.11 45 ++ 343.11 46 + 355.14 47++ 387.13 48 + 395.03 49 + 375.11 50 ++ 372.13 51 ++ 371.40 52 ++ 412.4553 ++ 366.39 54 ++ 343.35 55 +++ 373.37 56 + 366.39 57 +++ 486.53 58 ++379.33 59 +++ 379.33 60 +++ 372.39 61 + 363.33 62 +++ 403.40 63 ++385.39 64 + 385.43 65 + 387.40 66 + 426.50 67 ++ 485.55 68 ++ 408.4269 + 378.39 70 + 334.34 71 + 319.33 72 +++ 431.45 73 + 410.27 74 +++370.41 75 + 340.39 76 ++ 296.33 77 + 281.32 78 ++ 356.38 79 ++ 326.3680 + 335.20 81 + 282.30 82 + 267.29 83 ++ 418.42 84 ++ 357.37 85 +342.36 86 + 351.41 87 + 388.39 88 + 329.32 89 + 413.44 90 ++ 418.42 91++ 388.39 92 + 344.34 93 ++ 329.32 94 + 370.41 95 + 356.39 96 ++ 435.4697 ++ 361.38 98 + 346.37 99 + 373.44 100 + 430.49 101 + 418.46 102 +++418.42 103 ++ 388.39 104 ++ 413.44 105 ++ 378.39 106 ++ 348.36 107 ++425.49 108 ++ 439.52 109 + 464.48 110 +++ 418.46 111 ++ 388.39 112 +++418.42 113 +++ 418.42 114 ++ 394.50 115 ++ 364.50 116 ++ 389.50 117 +296.33 118 ++ 325.37 119 + 390.83 120 ++ 328.37 121 ++ 354.41 122 +413.40 123 ++ 354.41 124 ++ 416.44 125 + 355.40 126 + 401.43 127 +372.45 128 ++ 389.40 129 + 389.40 130 + 388.42 131 ++ 431.44 132 +431.44 133 + 413.43 134 + 413.43 135 + 391.42 136 + 391.42 137 + 390.44138 + 433.46 139 ++ 480.55 140 + 449.55 141 +++ 435.52 142 +++ 421.49143 +++ 435.52 144 +++ 507.63 145 ++ 435.52 146 +++ 463.53 147 +++435.52 148 +++ 525.60 149 +++ 425.48 150 ++ 467.52 151 +++ 525.60 152 ++HCl-salt: 461.94 153 ++ 467.52 154 ++ 419.43 155 ++ 425.53 156 +++425.53 157 +++ 453.54 158 +++ 439.51 159 +++ 431.48 160 ++ 507.58 161 ++521.61 162 +++ 425.53 163 + 421.54 164 + 417.46 165 +++ 439.55 166 ++378.40 167 ++ 426.47

Pharmacokinetic Studies

Compounds disclosed herein may be evaluated in pharmacokinetic assaysand models to determine absorption, distribution, metabolism, andexcretion parameters. The choice and tailoring of in vitro and ex vivoassays and in vivo models will vary according to the route ofadministration/formulation, indication under study, properties of testcompounds, etc., as well as according to such factors as costs,availability of technology and resources, etc. Such parameters are wellknown in the fields of pharmacology and drug development. It is withinthe capacity of one skilled in the art to design and carry out, suchwork, or to outsource it to a capable third party.

Pharmacokinetic Evaluation in Mice

Several compounds disclosed herein were evaluated in a standard murinepharmacokinetic model. Compounds were selected that exhibited reasonablesolubility and metabolic stability, and good predicted blood brainbarrier penetration, based on low molecular weight, a low number ofhydrogen bond donors, logD within a range of 2-4, and low polar surfacearea.

Compounds were dissolved in either 5% DMSO, 40% PEG400, and 55% saline(pH=8) or % DMSO, 40% PEG400, and 55% (20% HP-β-CD in deionized water;pH=8) to yield a nominal concentration of 2 mg/mL for intravenousadministration. Compounds were administered via a single intravenous(IV) injection in CL57 BL/6 mice at 10 mg/kg in DMSO/PEG400 solution.Three mice in each group were used for blood and brain collection ateach time point. Blood samples (300 μL) were collected via theretro-orbital vein predose and at 5 min, 0.25, 0.50, 1, 2, 4, 6, 8, and24 hours postdose. Blood samples were placed into tubes containingsodium heparin and centrifuged under refrigerated conditions at 8000 rpmfor 6 minutes to separate plasma from the samples. The brain of eachanimal was collected after the final blood collection. The whole tissuewas harvested, excised and rinsed by saline, dried by filter paper, andthen placed into one tube per tissue per animal. All samples were storedat −20° C. until bioanalysis.

Compound concentrations in plasma and brain homogenate were determinedusing a high performance liquid chromatography/mass spectrometry(HPLC/MS/MS) method (Agilent 1100 series HPLC, AB Inc. API4000triple-quadrupole with an ESI interface and Analyst 1.4 software).

Results in the form of area under the time-versus-concentration curve(AUC) are given below in Table 5. Additional compounds disclosed hereincan be tested according to this method and are expected to exhibitsimilar results.

TABLE 5 AUC Plasma AUC Brain + indicates ≧1500 + indicates ≧500 Ex. −indicates <1500 − indicates <500 1 + + 4 − − 9 + + 17 + − 18 + − 32 + +

Animal Models of Efficacy

Compounds disclosed herein may be tested in any number of well-known andpublicly available animal models of efficacy for diseases in which MLK3inhibition may play a therapeutic role. It is within the capacity of oneskilled in the art to select and tailor such a model.

Testing of Compounds for Efficacy in Established HIV-1-Encephalitic(HIVE) Mouse Model

For example, compounds disclosed herein can be ranked for in vivoefficacy in a mouse model relevant to NeuroAids (D. Eggert, The Journalof Immunology, in press, November 2009.) Test compounds selected may beprioritized based on MLK3 potency and favorable exposure in the brain,but this is not an absolute requirement. Four-week-old maleCB-17/IcrCrl-SCIDbr (CB17/SCID) mice may be purchased from Charles RiverLaboratory. HIV-1ADA-infected MDM (1.5×10⁵ cells infected at an MOI of0.1 in 5 ml) is stereotactically injected intracranially after 1 day ofviral infection and referred to as HIVE mice. The test compound is thenadministered i.p. daily for 7 days at doses 0.5, 1.0, 1.5, 5.0, and 15.0mg/kg/d (where, e.g., n=4 mice/treatment group). Vehicle only serves asthe control. CB17/SCID mice receive intracranial (i.c.) injections ofmedia (sham-operated) and serve as additional controls Animals aretreated with vehicle or test compound (i.e., a compound as disclosedherein) starting 1 d post-i.c. injection and for 7 d after MDMinjections and test compound treatments. Dosing parameters, number pergroup, etc. may be varied as needed, and such variations are within theskill of one skilled in the art.

Histopathology and Image Analysis

Brain tissue is collected at necropsy, fixed in 4% phosphate-bufferedparaformaldehyde, and embedded in paraffin. Paraffin blocks are cutuntil the injection site of the human MDM is identified. HIV-1 p24 Ag(cloneKal-1; Dako, Carpinteria, Calif.) is used to test forvirus-infected human MDM. For each mouse, 30-100 serial (5-mm-thick)sections are cut from the injection site and three to seven sections (10sections apart) analyzed. Abs to vimentin intermediate filaments (cloneVIM 3B4; Boehringer Mannheim, Indianapolis, Ind.) are used for detectionof human cells in mouse brains. Mouse microglia are detected by Abs toIba-1 (WAKO, Osaka, Japan), and astrocytes are detected by Abs for glialfibrillaryacidic protein ([GFAP] Dako). NeuN, MAP-2 (both from ChemiconInternational), and H chain (200 kDa) neurofilaments (Dako) are used fordetection of neurons. All sections are counterstained with Mayer'shematoxylin. The numbers of human MDM and HIV-1 p24 Ag-positive cellsare counted with a Nikon Microphot-FXA microscope. All obtained imagesare imported into Image-Pro Plus, v. 4.0 (Media Cybernetics, SilverSpring, Md.) for quantifying area (%) of GFAP, Iba-1, MAP-2, and NeuNpositive staining Efficacious MLK inhibitors will exhibit adose-dependent reduction in microgliosis and restoration of normalsynaptic architecture relative to control animals. Compounds disclosedherein can be tested according to this method and are expected toexhibit similar results.

From the foregoing description, one skilled in the art can easilyascertain the essential characteristics of this invention, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications of the invention to adapt it to various usages andconditions.

1-3. (canceled)
 4. A compound having a structure chosen from Formula VI,Formula VII, Formula VIII, or Formula IX:

or a salt thereof, wherein: Y₃ is chosen from a bond, lower alkyl, lowercarboxyl, and lower heteroalkyl; Y₄ is chosen from C(O), CHF, CF₂,—(CH₂)_(m)—, —(CH₂)_(m)O—, and —(CH₂)_(m)N—; m is an integer from 0 to1; R₂ is chosen from lower cycloalkyl, lower heterocycloalkyl, loweraryl, and lower heteroaryl, any of which are optionally substituted withone or more substituents chosen from halogen, hydroxy, cyano, C₁-C₄alkyl, C₂-C₄ alkenyl, C₁-C₄ alkynyl, C₁-C₄ haloalkyl, C₁-C₃ alkoxy,C₁-C₃ haloalkoxy, lower amino, lower amido, lower sulfonamido, and lowersulfonyl; R³ is chosen from lower cycloalkyl, lower heterocycloalkyl,lower aryl, and lower heteroaryl, any of which are optionallysubstituted with one or more substituents chosen from halogen, hydroxy,lower amino, lower amido, lower phenylamido, lower phenylalkylamido,lower heterocycloalkyl, lower alkylheterocycloalkyl, C₁-C₃ alkoxy andC₁-C₃ alkyl; R₁₄ is chosen from null, lower cycloalkyl, lowerheterocycloalkyl, phenyl, and lower heteroaryl, any of which may beoptionally substituted, wherein when the compound is chosen from FormulaVI, Y₃ is a bond, m is 0, R₁₄ is null and R₃ is indolyl, then R₂ is not3,4,5-trimethoxyphenyl, 4-hydroxyphenyl, or 4-morpholinophenyl; andwherein when the compound is chosen from Formula VI, Y₃ is a bond, m is0, R₁₄ is null, and R₃ is cyclopentyl, then R₂ is not3,4,5-trimethoxyphenyl or pyridinyl.
 5. (canceled)
 6. The compound asrecited in claim 4, wherein Y₃ is chosen from a bond, C(O), and loweralkyl.
 7. The compound as recited in claim 6, wherein R₃ is chosen fromlower cycloalkyl, phenyl, and lower heteroaryl, any of which may beoptionally substituted.
 8. (canceled)
 9. The compound as recited inclaim 7, wherein R₂ is chosen from phenyl, 6-membered monocyclicheteroaryl, and 5/6-fused bicyclic heteroaryl, any of which may beoptionally substituted.
 10. The compound as recited in claim 9, whereinR₂ is chosen from phenyl, pyridinyl, pyrimidinyl, and indolyl, any ofwhich may be optionally substituted.
 11. The compound as recited inclaim 10, wherein R₂ is substituted with one or more substituents chosenfrom halogen, hydroxy, lower amino, C₁-C₃ alkoxy and C₁-C₃ alkyl. 12.The compound as recited in claim 10, wherein R₂ is substituted with oneor more substituents chosen from fluorine, hydroxy, NH₂, NH(CH₃),N(CH₃)₂, methoxy, and methyl. 13-15. (canceled)
 16. The compound asrecited in claim 4, wherein the moiety R₁₄—Y₄—R₂ is chosen from

wherein u is an integer from 0 to 3; Y₄ is chosen from C(O),—(CH₂)_(m)—, —(CH₂)_(m)O—, and —(CH₂)_(m)N—; each R₁₅ is independentlychosen from halogen, hydroxy, cyano, C₁-C₄ alkyl, C₂-C₄ alkenyl, C₂-C₄alkynyl, C₁-C₄ haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, lower amino,lower amido, lower sulfonamido, and lower sulfonyl.
 17. (canceled) 18.The compound as recited in claim 16, wherein Y₄ is —(CH₂)_(m)—, M is 0;R₁₄ is null; u is an integer from 1 to 3; and each R₁₅ is independentlychosen from fluorine, hydroxy, NH₂, NH(CH₃), N(CH₃)₂, NS(O)₂CH₃,methoxy, and methyl.
 19. The compound as recited in claim 16, wherein Y₄is —(CH₂)_(m)—; M is 1; u is an integer from 0 to 2; and if present,each R₁₅ is independently chosen from fluorine, hydroxy, NH₂, NH(CH₃),N(CH₃)₂, NS(O)₂CH₃, methoxy, and methyl. 20-22. (canceled)
 23. Thecompound as recited in claim 16, wherein Y₃ is chosen from a bond andmethyl and wherein R₁₄ is chosen from null and optionally substitutedlower heterocycloalkyl. 24-26. (canceled)
 27. The compound as recited inclaim 16, wherein R₃ is chosen from benzothiazolyl, pyrrolopyridinyl,indanyl, cyclopropyl, cyclopentyl, phenyl, pyridinyl, pyrimidinyl, andindolyl, any of which may be optionally substituted. 28-29. (canceled)30. The compound as recited in claim 27, wherein R₃ is substituted withone or more substituents chosen from fluorine, chlorine, hydroxy, NH₂,NH(CH₃), N(CH₃)₂, C(O)NH₂, C(O)NHCH₃, morpholino, piperazinyl,methylpiperazinyl, acetamido, methylacetamido, methylpropionamido,phenylacetamidomethylene, benzamidomethylene,phenylpropanamidomethylene, methoxy and methyl.
 31. The compound asrecited in claim 4, wherein Y₃ is a bond; Y₄ is chosen from C(O), CH₂,CHF, and CF₂; R₂ is chosen from phenyl and 6-membered monocyclicheteroaryl, either of which may be optionally substituted; R₃ isoptionally substituted 5/6-fused bicyclic heteroaryl; R₁₄ is optionallysubstituted monocyclic heterocycloalkyl.
 32. (canceled)
 33. (canceled)34. The compound as recited in claim 31, wherein R₁₄ is optionallysubstituted piperazinyl. 35-42. (canceled)
 43. A pharmaceuticalcomposition comprising a compound as recited in claim 4 together with apharmaceutically acceptable carrier. 44-57. (canceled)
 58. A method oftreatment of a neuropsychiatric disorder in a patient, comprising thestep of administering to said patient a therapeutically effective amountof a compound as recited in claim
 4. 59. (canceled)
 60. The method asrecited in claim 58, wherein said neuropsychiatric disorder is chosenfrom depression, bipolar disorder, and post-traumatic stress disorder(PTSD).
 61. The method as recited in claim 58 wherein saidneuropsychiatric disorder is a traumatic brain injury.
 62. (canceled)63. The method as recited in claim 58 wherein said neuropsychiatricdisorder is chosen from Alzheimer's Disease (AD), Parkinson's Disease,and HIV associated neurocognitive disorder (HAND).
 64. The method asrecited in claim 58 wherein said neuropsychiatric disorder is aneurologic disorder of hearing or vision.
 65. The method as recited inclaim 64 wherein said neuropsychiatric disorder is chosen fromototoxicity, hearing loss, acute injury to the inner ear, acoustictrauma, and injury resulting from blast noise. 66-71. (canceled)